10.1186/2194-0517-2-14

Evaluation of silk sericin as a biomaterial: in vitro growth of human corneal limbal epithelial cells on Bombyx mori sericin membranes

HTML PDF
  • Traian V Chirila*1,
  • Shuko Suzuki2,
  • Laura J Bray3,
  • Nigel L Barnett4,
  • Damien G Harkin5,
  1. Queensland Eye Institute, South Brisbane, Queensland, 4101, AU Faculty of Science and Engineering, Queensland University of Technology, Brisbane, Queensland, 4001, AU Faculty of Health Sciences, The University of Queensland, Herston, Queensland, 4029, AU Australian Institute of Bioengineering & Nanotechnology, The University of Queensland, St Lucia, Queensland, 4072, AU Faculty of Science, The University of Western Australia, Crawley, Western Australia, 6009, AU
  2. Queensland Eye Institute, South Brisbane, Queensland, 4101, AU
  3. Queensland Eye Institute, South Brisbane, Queensland, 4101, AU Faculty of Health, Queensland University of Technology, Brisbane, Queensland, 4001, AU Max Bergmann Center of Biomaterials, Leibniz Institute for Polymer Research, Dresden, Saxony, 01069, DE
  4. Queensland Eye Institute, South Brisbane, Queensland, 4101, AU Faculty of Health, Queensland University of Technology, Brisbane, Queensland, 4001, AU UQ Centre for Clinical Research, The University of Queensland, Herston, Queensland, 4029, AU
  5. Queensland Eye Institute, South Brisbane, Queensland, 4101, AU Faculty of Health, Queensland University of Technology, Brisbane, Queensland, 4001, AU Institute of Health and Biomedical Innovation, Kelvin Grove, Queensland, 4059, AU
Cover Image

Published in Issue 2013-11-28

How to Cite

Chirila, T. V., Suzuki, S., Bray, L. J., Barnett, N. L., & Harkin, D. G. (2013). Evaluation of silk sericin as a biomaterial: in vitro growth of human corneal limbal epithelial cells on Bombyx mori sericin membranes. Progress in Biomaterials, 2(1 (December 2013). https://doi.org/10.1186/2194-0517-2-14
Crossref
Scopus
Google Scholar
Europe PMC

Abstract

Abstract Sericin and fibroin are the two major proteins in the silk fibre produced by the domesticated silkworm, Bombyx mori . Fibroin has been extensively investigated as a biomaterial. We have previously shown that fibroin can function successfully as a substratum for growing cells of the eye. Sericin has been so far neglected as a biomaterial because of suspected allergenic activity. However, this misconception has now been dispelled, and sericin’s biocompatibility is currently indisputable. Aiming at promoting sericin as a possible substratum for the growth of corneal cells in order to make tissue-engineered constructs for the restoration of the ocular surface, in this study we investigated the attachment and growth in vitro of human corneal limbal epithelial cells (HLECs) on sericin-based membranes. Sericin was isolated and regenerated from the silkworm cocoons by an aqueous procedure, manufactured into membranes, and characterized (mechanical properties, structural analysis, contact angles). Primary cell cultures from two donors were established in serum-supplemented media in the presence of murine feeder cells. Membranes made of sericin and fibroin-sericin blends were assessed in vitro as substrata for HLECs in a serum-free medium, in a cell attachment assay and in a 3-day cell growth experiment. While the mechanical characteristics of sericin were found to be inferior to those of fibroin, its ability to enhance the attachment of HLECs was significantly superior to fibroin, as revealed by the PicoGreen ® assay. Evidence was also obtained that cells can grow and differentiate on these substrata.

Keywords

  • Silk,
  • Silk proteins,
  • Sericin,
  • Corneal limbal epithelial cells,
  • Cell attachment
HTML PDF

References

  1. Altman et al. (2003) Silk-based biomaterials (pp. 401-416) https://doi.org/10.1016/S0142-9612(02)00353-8
  2. Aramwit et al. (2009a) Monitoring of inflammatory mediators induced by silk sericin (pp. 556-561) https://doi.org/10.1016/j.jbiosc.2008.12.012
  3. Aramwit et al. (2009b) The effect of sericin with variable amino-acid content from different silk strains on the production of collagen and nitric oxide (pp. 1295-1306) https://doi.org/10.1163/156856209X453006
  4. Aramwit et al. (2010) The effect of sericin from various extraction methods on cell viability and collagen production (pp. 2200-2211) https://doi.org/10.3390/ijms11052200
  5. Bray et al. (2011) Human corneal epithelial equivalents constructed on Bombyx mori silk fibroin membranes (pp. 5086-5091) https://doi.org/10.1016/j.biomaterials.2011.03.068
  6. Bray et al. (2012) A dual-layer silk fibroin scaffold for reconstructing the human corneal limbus (pp. 3529-3538) https://doi.org/10.1016/j.biomaterials.2012.01.045
  7. Bray et al. (2013) Incorporation of exogenous RGD peptide and inter-species blending as strategies for enhancing human corneal limbal epithelial cell growth on Bombyx mori silk fibroin membranes (pp. 74-88) https://doi.org/10.3390/jfb4020074
  8. Brown and Coleman (1957) Severe immediate reactions to biologicals caused by silk allergy (pp. 2178-2180) https://doi.org/10.1001/jama.1957.02980350036008
  9. Bryant (1948) The sericin fractions of raw silk
  10. Celedón et al. (2001) Sensitization to silk and childhood asthma in rural China https://doi.org/10.1542/peds.107.5.e80
  11. Chirila et al. (2007) Silk as substratum for cell attachment and proliferation (pp. 1549-1552) https://doi.org/10.4028/www.scientific.net/MSF.561-565.1549
  12. Chirila et al. (2008) Bombyx mori silk fibroin membranes as potential substrata for epithelial constructs used in the management of ocular surface disorders (pp. 1203-1211) https://doi.org/10.1089/ten.tea.2007.0224
  13. Chirila et al. (2010) Reconstruction of the ocular surface using biomaterials (pp. 213-242) Woodhead/CRC https://doi.org/10.1533/9781845697433.1.213
  14. Clarke and Meyer (1923) A case of hypersensitiveness to silk (pp. 11-12) https://doi.org/10.1001/jama.1923.02640280013004
  15. Unknown (1785) Observations sur la dissolution du vernis de la soie [Observations on the dissolution of the varnish of the silk] 27(II) (pp. 95-107)
  16. Dewair et al. (1985) Use of immunoblot technique for detection of human IgE and IgG antibodies to individual silk proteins (pp. 537-542) https://doi.org/10.1016/0091-6749(85)90772-9
  17. Djerassi et al. (1960) Naturally occurring oxygen heterocycles: IX. Isolation and characterization of genipin (pp. 2174-2177) https://doi.org/10.1021/jo01082a022
  18. Fedič et al. (2002) The silk of Lepidoptera (pp. 1-15)
  19. Figley and Parkhurst (1933) Silk sensitivity (pp. 60-69) https://doi.org/10.1016/S0021-8707(33)90171-9
  20. Friedman et al. (1957) Severe allergic reaction caused by silk as a contaminant in typhoid-parathyphoid vaccine (pp. 489-493) https://doi.org/10.1016/0021-8707(57)90078-3
  21. Gamo (1982) Genetic variants of the Bombyx mori silkworm encoding sericin proteins of different lengths (pp. 165-177) https://doi.org/10.1007/BF00484944
  22. Gamo et al. (1977) Polypeptides of fibroin and sericin secreted from the different sections of the silk gland (pp. 285-295) https://doi.org/10.1016/0020-1790(77)90026-9
  23. George et al. (2013) Effect of sterilization method on the properties of Bombyx mori silk fibroin films (pp. 668-674) https://doi.org/10.1016/j.msec.2012.10.016
  24. Ghassemifar et al. (2010) Advancing towards a tissue-engineered tympanic membrane: silk fibroin as a substratum for growing human eardrum keratinocytes (pp. 591-606) https://doi.org/10.1177/0885328209104289
  25. Gil et al. (2013) Functionalized silk biomaterials for wound healing (pp. 206-217) https://doi.org/10.1002/adhm.201200192
  26. Grzelak (1995) Control of expression of silk protein genes (pp. 671-681) https://doi.org/10.1016/0305-0491(94)00215-G
  27. Häcki et al. (1982) Widseide: ein aggressives Inhalationsallergen [Wild silk: a strong inhalation allergen] (pp. 166-169) https://doi.org/10.1055/s-2008-1069892
  28. Hakimi et al. (2007) Spider and mulberry silkworm silks as compatible biomaterials (pp. 324-337) https://doi.org/10.1016/j.compositesb.2006.06.012
  29. Hakimi et al. (2010) Modulation of cell growth on exposure to silkworm and spider silk fibers (pp. 1366-1372)
  30. Hardy and Scheibel (2010) Composite materials based on silk proteins (pp. 1093-1115) https://doi.org/10.1016/j.progpolymsci.2010.04.005
  31. Harindranath et al. (1985) Prevalence of occupational asthma in silk filatures (pp. 511-515)
  32. Harkin and Chirila (2012) Silk fibroin in ocular surface reconstruction - what is its potential as a biomaterial in ophthalmics? (pp. 2145-2147) https://doi.org/10.4155/fmc.12.155
  33. Harkin et al. (2011) Silk fibroin in ocular tissue reconstruction (pp. 2445-2458) https://doi.org/10.1016/j.biomaterials.2010.12.041
  34. Hjortdal (1996) Regional elastic performance of the human cornea (pp. 931-942) https://doi.org/10.1016/0021-9290(95)00152-2
  35. Horbett et al. (1996) Cell culturing: surface aspects and considerations (pp. 351-445) Marcel Dekker Inc
  36. Hu et al. (2006) Determining beta-sheet crystallinity in fibrous proteins by thermal analysis and infrared spectroscopy (pp. 6161-6170) https://doi.org/10.1021/ma0610109
  37. Hu et al. (2011) Regulation of silk material structure by temperature-controlled water vapour annealing (pp. 1686-1696) https://doi.org/10.1021/bm200062a
  38. Inoue et al. (2000) 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 (pp. 40517-40528) https://doi.org/10.1074/jbc.M006897200
  39. Johansson et al. (1985) Nightly asthma caused by allergens in silk-filled bed quilts: clinical and immunologic studies (pp. 452-459) https://doi.org/10.1016/S0091-6749(85)80017-8
  40. Julien et al. (2005) Silk gland development and regulation of silk protein genes (pp. 369-384) Elsevier BV https://doi.org/10.1016/B0-44-451924-6/00022-3
  41. Kearns et al. (2008) Silk-based biomaterials for tissue engineering (pp. 1-19)
  42. Kikkawa (1953) Biochemical genetics of Bombyx mori (silkworm) (pp. 107-140) https://doi.org/10.1016/S0065-2660(08)60407-1
  43. Kino and Oshima (1979) Allergy to insects in Japan: II. The reaginic sensitivity to silkworm moth in patients with bronchial asthma (pp. 131-138) https://doi.org/10.1016/0091-6749(79)90047-2
  44. Kodama (1926) The preparation and physico-chemical properties of sericin (pp. 1208-1222) https://doi.org/10.1042/bj0201208
  45. Kondo (1921) Research on some properties of sericin [in Japanese] (pp. 1054-1065)
  46. Kundu et al. (2008) Natural protective glue protein, sericin bioengineered by silkworms: potential for biomedical and biotechnological applications (pp. 998-1012) https://doi.org/10.1016/j.progpolymsci.2008.08.002
  47. Kwan et al. (2010) Development of tissue-engineered membranes for the culture and transplantation of retinal pigment epithelial cells (pp. 390-408) Woodhead/CRC https://doi.org/10.1533/9781845697433.2.390
  48. Lee (2004) Silk sericin retards the crystallization of silk fibroin (pp. 1792-1796) https://doi.org/10.1002/marc.200400333
  49. Lucas et al. (1958) The silk fibroins (pp. 107-242) https://doi.org/10.1016/S0065-3233(08)60599-9
  50. Madden et al. (2011) Human corneal endothelial cell growth on a silk fibroin membrane (pp. 4076-4084) https://doi.org/10.1016/j.biomaterials.2010.12.034
  51. Mase et al. (2006) A new silkworm race for sericin production, “SERICIN HOPE” and its product, “VIRGIN SERICIN” (pp. 85-88)
  52. Michaille et al. (1986) A single gene produces multiple sericin messenger RNAs in the silk gland of Bombyx mori (pp. 1165-1173) https://doi.org/10.1016/S0300-9084(86)80060-8
  53. Michaille et al. (1989) The expression of five middle silk gland specific genes is territorially regulated during the larval development of Bombyx mori (pp. 19-27) https://doi.org/10.1016/0020-1790(89)90005-X
  54. Minoura et al. (1990) Physico-chemical properties of silk fibroin membrane as a biomaterial (pp. 430-434) https://doi.org/10.1016/0142-9612(90)90100-5
  55. Minoura et al. (1995a) Attachment and growth of fibroblast cells on silk fibroin (pp. 511-516) https://doi.org/10.1006/bbrc.1995.1368
  56. Minoura et al. (1995b) Attachment and growth of cultured fibroblast cells on silk protein matrices (pp. 1215-1221) https://doi.org/10.1002/jbm.820291008
  57. Mondal et al. (2007) The silk proteins, sericin and fibroin in silkworm, Bombyx mori Linn., - a review (pp. 63-76)
  58. Mosher (1934) The sericin fractions of silk (pp. 43-44)
  59. Motta et al. (2011) Stabilization of Bombyx mori silk fibroin/sericin films by crosslinking with PEG-DE 600 and genipin (pp. 130-143) https://doi.org/10.1177/0883911511400251
  60. Murphy and Kaplan (2009) Biomedical applications of chemically-modified silk fibroin (pp. 6443-6450) https://doi.org/10.1039/b905802h
  61. Nagura et al. (2001) Structures and physical properties of crosslinked sericin membranes (pp. 149-153)
  62. Nakazawa and Umegae (1990) Sericulturist’s lung disease: hypersensitivity pneumonitis related to silk production (pp. 233-234) https://doi.org/10.1136/thx.45.3.233
  63. Numata and Kaplan (2010) Silk-based delivery systems of bioactive molecules (pp. 1497-1508) https://doi.org/10.1016/j.addr.2010.03.009
  64. Panilaitis et al. (2003) Macrophage responses to silk (pp. 3079-3085) https://doi.org/10.1016/S0142-9612(03)00158-3
  65. Pritchard and Kaplan (2011) Silk fibroin biomaterials for controlled release drug delivery (pp. 797-811) https://doi.org/10.1517/17425247.2011.568936
  66. Roard (1808) Mémoire sur le décreusage de la soie [Dissertation on the degumming of the silk] (pp. 44-58)
  67. Rutherford and Harris (1940) Concerning the existence of fractions of the sericin in raw silk (pp. 221-228) https://doi.org/10.1177/004051754001000601
  68. Salthouse et al. (1977) Comparative tissue response to six suture materials in rabbit cornea, sclera, and ocular muscle (pp. 224-233) https://doi.org/10.1016/0002-9394(77)90856-X
  69. Sehnal et al. (2011) Biotechnologies based on silk (pp. 211-224) Springer https://doi.org/10.1007/978-90-481-9641-8_11
  70. Shadforth et al. (2012) The cultivation of human retinal pigment epithelial cells on Bombyx mori silk fibroin (pp. 4110-4117) https://doi.org/10.1016/j.biomaterials.2012.02.040
  71. Shelton and Johnson (1925) Research on proteins: VII. The preparation of the protein “sericin” from silk (pp. 412-418) https://doi.org/10.1021/ja01679a020
  72. Soong and Kenyon (1984) Adverse reactions to virgin silk sutures in cataract surgery (pp. 479-483) https://doi.org/10.1016/S0161-6420(84)34273-7
  73. Sprague (1975) The Bombyx mori silk proteins characterization of large polypeptides (pp. 925-931) https://doi.org/10.1021/bi00676a008
  74. Suzuki et al. (1995) Causative allergens of allergic rhinitis in Japan with special reference to silkworm moth allergen (pp. 23-27) https://doi.org/10.1111/j.1398-9995.1995.tb02479.x
  75. Takasu et al. (2002) Isolation of three main sericin components from the cocoon of the silkworm, Bombyx mori (pp. 2715-2718) https://doi.org/10.1271/bbb.66.2715
  76. Tashiro and Otsuki (1970) Studies on the posterior silk gland of the silkworm Bombyx mori (pp. 1-16) https://doi.org/10.1083/jcb.46.1.1
  77. Taub (1930) Allergy due to silk (pp. 539-541) https://doi.org/10.1016/S0021-8707(30)90208-0
  78. Terada et al. (2002) Sericin, a protein derived from silkworms, accelerates the proliferation of several mammalian cell lines including a hybridoma (pp. 3-12) https://doi.org/10.1023/A:1023993400608
  79. Teramoto and Miyazawa (2005) Molecular orientation behaviour of silk sericin as revealed by ATR infrared spectroscopy (pp. 2049-2057) https://doi.org/10.1021/bm0500547
  80. Teramoto et al. (2005) Preparation of elastic sericin hydrogel (pp. 845-847) https://doi.org/10.1271/bbb.69.845
  81. Teramoto et al. (2006) Native structure and degradation pattern of silk sericin studied by 13C NMR spectroscopy (pp. 6-8) https://doi.org/10.1021/ma0521147
  82. Teramoto et al. (2008) Preparation of gel film from Bombyx mori silk sericin and its characterization as a wound dressing (pp. 3189-3196) https://doi.org/10.1271/bbb.80375
  83. Tokutake (1980) Isolation of smallest component of silk protein (pp. 413-417) https://doi.org/10.1042/bj1870413
  84. Tsubouchi et al. (2005) Sericin enhances attachment of cultured human skin fibroblasts (pp. 403-405) https://doi.org/10.1271/bbb.69.403
  85. Vepari and Kaplan (2007) Silk as a biomaterial (pp. 991-1007) https://doi.org/10.1016/j.progpolymsci.2007.05.013
  86. Wang et al. (2006) Stem cell-based tissue engineering with silk biomaterials (pp. 6064-6082) https://doi.org/10.1016/j.biomaterials.2006.07.008
  87. Wang et al. (2009) Silk proteins - biomaterials and engineering (pp. 939-959) Wiley
  88. Wenk et al. (2011) Silk fibroin as a vehicle for drug delivery applications (pp. 128-141) https://doi.org/10.1016/j.jconrel.2010.11.007
  89. Xie et al. (2007) Preparation of sericin film and its cytocompatibility (pp. 241-244) https://doi.org/10.4028/www.scientific.net/KEM.342-343.241
  90. Zeng et al. (2001) A comparison of biomechanical properties between human and porcine cornea (pp. 533-537) https://doi.org/10.1016/S0021-9290(00)00219-0
  91. Zhaoming et al. (1990) Silk-induced asthma in children: a report of 64 cases (pp. 365-378)
  92. Zhaoming et al. (1996) Partial characterization of the silk allergens in mulberry silk extract (pp. 237-241)