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Bioartificial Sponges for Auricular Cartilage Engineering

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Advances in Bionanomaterials II (BIONAM 2019 2019)

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Abstract

Auricle reconstruction due to congenital, post-infective or post-traumatic defects represents a challenging procedure in the field of aesthetic and reconstructive surgery due to the highly complex three-dimensional anatomy of the outer ear. Tissue engineering aims to provide alternatives to overcome the shortcomings of standard surgical reconstructive procedure. In the present study, poly(vinyl alcohol)/gelatin (PVA/G) sponges at different weight ratios were produced via emulsion and freeze-drying, and crosslinked by exposure to glutaraldehyde vapors. PVA/G sponges gave rise to highly porous, water stable and hydrophilic scaffolds. Characterization of PVA/G sponges showed round-shaped interconnected pores, high swelling capacity (>200%) and viscoelastic mechanical behavior. The PVA/G 70/30 (w/w) scaffold was selected for in vitro biological studies. Bone marrow derived human mesenchymal stromal cells (hMSCs) were used and differentiated towards chondrogenic lineage under different culture conditions: 1) commercial versus handmade differentiation medium; 2) undifferentiated versus pre-differentiated hMSC seeding; and 3) static versus dynamic culture [i.e. ultrasound (US) or bioreactor stimulation]. Histological results highlighted intense glycosaminoglycan, glycoprotein and collagen syntheses after three weeks, mostly using the commercial medium, whereas round morphology was observed in pre-differentiated cells. In static culture, immunohistochemistry for chondrogenic markers revealed an early differentiation stage, characterized by the expression of Sox-9 and collagen type I fibers. The application of US on cell/scaffold constructs increased extracellular matrix deposition and resulted in 30% higher collagen type II expression at the gene level. Bioreactor culture induced collagen type II, aggrecan and elastin formation. This study demonstrated that 70/30 PVA/G sponge is a suitable candidate for auricle reconstruction.

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References

  1. Kamil, S.H., Vacanti, M.P., Vacanti, C.A., Eavey, R.D.: Microtia chondrocytes as a donor source for tissue-engineered cartilage. Laryngoscope 114, 2187–2190 (2004)

    Article  Google Scholar 

  2. Han, S.E., Lim, S.Y., Pyon, J.K., Bang, S.I., Mun, G.H., Oh, K.S.: Aesthetic auricular reconstruction with autologous rib cartilage grafts in adult microtia patients. J. Plast. Reconstr. Aesthetic Surg. 68, 1085–1094 (2015)

    Article  Google Scholar 

  3. Jessop, Z.M., Javed, M., Otto, I.A., Combellack, E.J., Morgan, S., Breugem, C.C., Archer, C.W., Khan, I.M., Lineaweaver, W.C., Kon, M., et al.: Combining regenerative medicine strategies to provide durable reconstructive options: auricular cartilage tissue engineering. Stem Cell Res. Ther. 7, 19 (2016)

    Article  Google Scholar 

  4. Griffin, M.F., Premakumar, Y., Seifalian, A.M., Szarko, M., Butler, P.E.M.: Biomechanical characterisation of the human auricular cartilages; implications for tissue engineering. Ann. Biomed. Eng. 44, 3460–3467 (2016)

    Article  Google Scholar 

  5. Thorne, C.H., Brecht, L.E., Bradley, J.P., Levine, J.P., Hammerschlag, P., Longaker, M.T.: Auricular reconstruction: Indications for autogenous and prosthetic techniques. Plastic Reconstr. Surg. 107, 1241–1252 (2001)

    Article  Google Scholar 

  6. Ciorba, A., Martini, A.: Tissue engineering and cartilage regeneration for auricular reconstruction. Int. J. Pediatr. Otorhinolaryngol. 70, 1507–1515 (2006)

    Article  Google Scholar 

  7. Cao, Y., Vacanti, J.P., Paige, K.T., Upton, J., Vacanti, C.A.: Transplantation of chondrocytes utilizing a polymer-cell construct to produce tissue-engineered cartilage in the shape of a human ear. Plastic Reconstr. Surg. 100, 297–302 (1997)

    Article  Google Scholar 

  8. Shieh, S.J., Terada, S., Vacanti, J.P.: Tissue engineering auricular reconstruction: in vitro and in vivo studies. Biomaterials 25, 1545–1557 (2004)

    Article  Google Scholar 

  9. Zhou, G., Jiang, H., Yin, Z., Liu, Y., Zhang, Q., Zhang, C., Pan, B., Zhou, J., Zhou, X., Sun, H., et al.: In vitro regeneration of patient-specific ear-shaped cartilage and its first clinical application for auricular reconstruction. EBioMedicine 28, 287–302 (2018)

    Article  Google Scholar 

  10. Schultz, T.W., Geneser, F.: Textbook of Histology. Transactions of the American Microscopical Society (1987)

    Google Scholar 

  11. Lai, C.H., Chen, S.C., Chiu, L.H., Yang, C.B., Tsai, Y.H., Zuo, C.S., Chang, W.H.S., Lai, W.F.: Effects of low-intensity pulsed ultrasound, dexamethasone/TGF-β1 and/or BMP-2 on the transcriptional expression of genes in human mesenchymal stem cells: chondrogenic vs. osteogenic differentiation. Ultrasound in Med. Biol. 36, 1022–1033 (2010)

    Article  Google Scholar 

  12. Murakami, W.T., Wong, L.W., Davidson, T.M.: Applications of the biomechanical behavior of cartilage to nasal septoplastic surgery. Laryngoscope 92, 300–309 (1982)

    Article  Google Scholar 

  13. Van Osch, G.J.V.M., Van Den Berg, W.B., Hunziker, E.B., Häuselmann, H.J.: Differential effects of IGF-1 and TGFβ-2 on the assembly of proteoglycans in pericellular and territorial matrix by cultured bovine articular chondrocytes. Osteoarthritis Cartilage 6, 187–195 (1998)

    Article  Google Scholar 

  14. Ackert, J.E., Maximow, A.A., Bloom, W.: A Textbook of Histology. Transactions of the American Microscopical Society (1942)

    Google Scholar 

  15. Gosline, J., Lillie, M., Carrington, E., Guerette, P., Ortlepp, C., Savage, K.: Elastic proteins: biological roles and mechanical properties. Philos. Trans. R. Soc. B: Biol. Sci. 357, 121–132 (2002)

    Article  Google Scholar 

  16. Lotz, M., Loeser, R.F.: Effects of aging on articular cartilage homeostasis. Bone 51, 241–248 (2012)

    Article  Google Scholar 

  17. Ross, M.H.P., Pawlina, W.: Histology a Text and Atlas with Correlated Cell and Molecular Biology (2014). ISBN 9780874216561

    Google Scholar 

  18. Xia, P., Wang, X., Qu, Y., Lin, Q., Cheng, K., Gao, M., Ren, S., Zhang, T., Li, X.: TGF-β1-induced chondrogenesis of bone marrow mesenchymal stem cells is promoted by low-intensity pulsed ultrasound through the integrin-mTOR signaling pathway. Stem Cell Res. Ther. 8, 281–292 (2017)

    Article  Google Scholar 

  19. Fung, Y.C., Skalak, R.: Biomechanics: Mechanical Properties of Living Tissues. Journal of Applied Mechanics (1982)

    Google Scholar 

  20. Urry, D.W., Hugel, T., Seitz, M., Gaub, H.E., Sheiba, L., Dea, J., Xu, J., Parker, T.: Elastin: a representative ideal protein elastomer. Philos. Trans. R. Soc. B: Biol. Sci. 357, 169–184 (2002)

    Article  Google Scholar 

  21. Milazzo, M., Jung, G.S., Danti, S., Buehler, M.J.: Wave propagation and energy dissipation in collagen molecules. ACS Biomater. Sci. Eng. 6, 1367–1374 (2020)

    Article  Google Scholar 

  22. Sherratt, M.J.: Tissue elasticity and the ageing elastic fibre. Age 31, 305–325 (2009)

    Article  Google Scholar 

  23. Nimeskern, L., Utomo, L., Lehtoviita, I., Fessel, G., Snedeker, J.G., van Osch, G.J.V.M., Müller, R., Stok, K.S.: Tissue composition regulates distinct viscoelastic responses in auricular and articular cartilage. J. Biomech. 49, 344–352 (2016)

    Article  Google Scholar 

  24. Riedler, K.L., Shokrani, A., Markarian, A., Fisher, L.M., Pepper, J.P.: Age-related histologic and biochemical changes in auricular and septal cartilage. Laryngoscope 127, 399–407 (2017)

    Article  Google Scholar 

  25. Vacanti, C.A., Vacanti, J.P.: Bone and cartilage reconstruction with tissue engineering approaches. Otolaryngol. Clin. North Am. 27, 263–276 (1994)

    Google Scholar 

  26. Rodriguez, A., Cao, Y.L., Ibarra, C., Pap, S., Vacanti, M., Eavey, R.D., Vacanti, C.A.: Characteristics of cartilage engineered from human pediatric auricular cartilage. Plastic Reconstr. Surg. 103, 1111–1119 (1999)

    Article  Google Scholar 

  27. Otto, I.A., Levato, R., Webb, W.R., Khan, I.M., Breugem, C.C., Malda, J.: Progenitor cells in auricular cartilage demonstrate cartilage-forming capacity in 3D hydrogel culture. Eur. Cells Mater 35, 132–150 (2018)

    Article  Google Scholar 

  28. Ciuffreda, M.C., Malpasso, G., Musarò, P., Turco, V., Gnecchi, M.: Protocols for in vitro differentiation of human mesenchymal stem cells into osteogenic, chondrogenic and adipogenic lineages. In: Mesenchymal Stem Cells, pp. 149–158. Springer (2016)

    Google Scholar 

  29. Pittenger, M.F., Mackay, A.M., Beck, S.C., Jaiswal, R.K., Douglas, R., Mosca, J.D., Moorman, M.A., Simonetti, D.W., Craig, S., Marshak, D.R.: Multilineage potential of adult human mesenchymal stem cells. Science 284, 143–147 (1999)

    Article  Google Scholar 

  30. Kusuhara, H., Isogai, N., Enjo, M., Otani, H., Ikada, Y., Jacquet, R., Lowder, E., Landis, W.J.: Tissue engineering a model for the human ear: assessment of size, shape, morphology, and gene expression following seeding of different chondrocytes. Wound Repair Regener. 17, 136–146 (2009)

    Article  Google Scholar 

  31. Milazzo, M., Contessi Negrini, N., Scialla, S., Marelli, B., Farè, S., Danti, S., Buehler, M.J.: Additive manufacturing approaches for hydroxyapatite-reinforced composites. Adv. Funct. Mater. 29, 1903055 (2019)

    Article  Google Scholar 

  32. Cascone, M.G., Lazzeri, L., Sparvoli, E., Scatena, M., Serino, L.P., Danti, S.: Morphological evaluation of bioartificial hydrogels as potential tissue engineering scaffolds. J. Mater. Sci. Mater. Med. 15, 1309–1313 (2004)

    Article  Google Scholar 

  33. Moscato, S., Mattii, L., D’Alessandro, D., Cascone, M.G., Lazzeri, L., Serino, L.P., Dolfi, A., Bernardini, N.: Interaction of human gingival fibroblasts with PVA/gelatine sponges. Micron 39, 569–579 (2008)

    Article  Google Scholar 

  34. Kamoun, E.A., Chen, X., Mohy Eldin, M.S., Kenawy, E.R.S.: Crosslinked poly(vinyl alcohol) hydrogels for wound dressing applications: a review of remarkably blended polymers. Arab. J. Chem. 8, 1–14 (2015)

    Article  Google Scholar 

  35. Lee, H.J., Choi, B.H., Min, B.H., Son, Y.S., Park, S.R.: Low-intensity ultrasound stimulation enhances chondrogenic differentiation in alginate culture of mesenchymal stem cells. Artif. Organs 30, 707–715 (2006)

    Article  Google Scholar 

  36. Bernardo, M.E., Fibbe, W.E.: Mesenchymal stromal cells and hematopoietic stem cell transplantation. Immunol. Lett. 168, 215–221 (2015)

    Article  Google Scholar 

  37. Parvizi, J., Wu, C.C., Lewallen, D.G., Greenleaf, J.F., Bolander, M.E.: Low-intensity ultrasound stimulates proteoglycan synthesis in rat chondrocytes by increasing aggrecan gene expression. J. Orthop. Res. 17, 488–494 (1999)

    Article  Google Scholar 

  38. Jonnalagadda, U.S., Hill, M., Messaoudi, W., Cook, R.B., Oreffo, R.O.C., Glynne-Jones, P., Tare, R.S.: Acoustically modulated biomechanical stimulation for human cartilage tissue engineering. Lab Chip 18, 473–485 (2018)

    Article  Google Scholar 

  39. Aliabouzar, M., Lee, S.J., Zhou, X., Zhang, G.L., Sarkar, K.: Effects of scaffold microstructure and low intensity pulsed ultrasound on chondrogenic differentiation of human mesenchymal stem cells. Biotechnol. Bioeng. 115, 495–506 (2018)

    Article  Google Scholar 

  40. Ricci, C., Danti, S.: 3D models of pancreatic ductal adenocarcinoma via tissue engineering. Methods Mol. Biol. 1882, 81–95 (2019)

    Article  Google Scholar 

  41. Mattei, G., Tirella, A., Gallone, G., Ahluwalia, A.: Viscoelastic characterisation of pig liver in unconfined compression. J. Biomech. 47, 2641–2646 (2014)

    Article  Google Scholar 

  42. Tirella, A., Mattei, G., Ahluwalia, A.: Strain rate viscoelastic analysis of soft and highly hydrated biomaterials. J. Biomed. Mater. Res. - Part A 102, 3352–3360 (2014)

    Article  Google Scholar 

  43. Barachini, S., Danti, S., Pacini, S., D’Alessandro, D., Carnicelli, V., Trombi, L., Moscato, S., Mannari, C., Cei, S., Petrini, M.: Plasticity of human dental pulp stromal cells with bioengineering platforms: a versatile tool for regenerative medicine. Micron 67, 155–168 (2014)

    Article  Google Scholar 

  44. Bajpai, V.K., Mistriotis, P., Andreadis, S.T.: Clonal multipotency and effect of long-term in vitro expansion on differentiation potential of human hair follicle derived mesenchymal stem cells. Stem Cell Res. 8, 74–84 (2012)

    Article  Google Scholar 

  45. Gurikov, P., Smirnova, I.: Non-conventional methods for gelation of alginate. Gels 4, 14 (2018)

    Article  Google Scholar 

  46. De la Ossa, J.G., Trombi, L., D’Alessandro, D., Coltelli, M.B., Serino, L.P., Pini, R., Lazzeri, A., Petrini, M., Danti, S.: Pore size distribution and blend composition affect in vitro prevascularized bone matrix formation on poly(vinyl alcohol)/gelatin sponges. Macromol. Mater. Eng. 302, 1700300 (2017)

    Article  Google Scholar 

  47. Chiellini, E., Cinelli, P., Fernandes, E.G., Kenawy, E.R.S., Lazzeri, A.: Gelatin-based blends and composites. Morphological and thermal mechanical characterization. Biomacromol 2, 806–811 (2001)

    Article  Google Scholar 

  48. Bigi, A., Cojazzi, G., Panzavolta, S., Rubini, K., Roveri, N.: Mechanical and thermal properties of gelatin films at different degrees of glutaraldehyde crosslinking. Biomaterials 22, 763–768 (2001)

    Article  Google Scholar 

  49. Alves, P.M.A., Carvalho, R.A., Moraes, I.C.F., Luciano, C.G., Bittante, A.M.Q.B., Sobral, P.J.A.: Development of films based on blends of gelatin and poly(vinyl alcohol) cross linked with glutaraldehyde. Food Hydrocolloids 25, 1751–1757 (2011)

    Article  Google Scholar 

  50. Gao, X., Tang, K., Liu, J., Zheng, X., Zhang, Y.: Compatibility and properties of biodegradable blend films with gelatin and poly (vinyl alcohol). J. Wuhan Univ. Technol.-Mater. Sci. Ed. 29, 351–356 (2014)

    Article  Google Scholar 

  51. Pawde, S.M., Deshmukh, K.: Characterization of polyvinyl alcohol/gelatin blend hydrogel films for biomedical applications. J. Appl. Polym. Sci. 109, 3431–3437 (2008)

    Article  Google Scholar 

  52. Mattei, G., Gruca, G., Rijnveld, N., Ahluwalia, A.: The nano-epsilon dot method for strain rate viscoelastic characterisation of soft biomaterials by spherical nano-indentation. J. Mech. Behav. Biomed. Mater. 50, 150–159 (2015)

    Article  Google Scholar 

  53. Cacopardo, L., Mattei, G., Ahluwalia, A.: A new load-controlled testing method for viscoelastic characterisation through stress-rate measurements. Materialia 9, 100552 (2020)

    Article  Google Scholar 

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Acknowledgements

AURICULAE Project, funded by Stem Cells & Life Foundation, Padova, Italy is greatly acknowledged. Dr. Delfo D’Alessandro (University of Pisa, Pisa, Italy), as well as Dr. Alessandra Fusco and Dr. Giovanna Donnarumma (University of Campania “Luigi Vanvitelli”, Naples, Italy) are thanked for their fundamental technical support to this work.

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Authors and Affiliations

  1. Research Center “E. Piaggio”, University of Pisa, 56122, Pisa, Italy

    Marta Feula, Ludovica Cacopardo, Arti Ahluwalia & Serena Danti

  2. The BioRobotics Institute, Scuola Superiore Sant’Anna, 56025, Pontedera, PI, Italy

    Mario Milazzo & Serena Danti

  3. Department of Civil and Industrial Engineering (DICI), University of Pisa, 56122, Pisa, Italy

    Giulia Giannone, Andrea Lazzeri & Serena Danti

  4. Research Unit of DICI-Pisa, Interuniversity Consortium for Materials Science and Technology (INSTM), 50121, Florence, Italy

    Bahareh Azimi & Luisa Trombi

  5. Department of Clinical and Experimental Medicine, University of Pisa, 56126, Pisa, Italy

    Stefania Moscato

  6. Department of Surgical, Medical, Molecular Pathology and Emergency Medicine, University of Pisa, 56126, Pisa, Italy

    Stefano Berrettini

  7. Institute for Technology Inspired Regenerative Medicine (MERLN), Complex Tissue Regeneration Department, Maastricht University, 6229ER, Maastricht, The Netherlands

    Carlos Mota

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  1. Marta Feula
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  2. Mario Milazzo
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Correspondence to Serena Danti .

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Editors and Affiliations

  1. Department of Pharmacy, University of Salerno, Fisciano, Italy

    Stefano Piotto

  2. Department of Industrial Engineering, University of Salerno, Fisciano, Italy

    Simona Concilio

  3. Department of Pharmacy, University of Salerno, Fisciano, Italy

    Lucia Sessa

  4. Department of Earth, Environmental and Physical Sciences, University of Siena, Siena, Italy

    Federico Rossi

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Feula, M. et al. (2020). Bioartificial Sponges for Auricular Cartilage Engineering. In: Piotto, S., Concilio, S., Sessa, L., Rossi, F. (eds) Advances in Bionanomaterials II. BIONAM 2019 2019. Lecture Notes in Bioengineering. Springer, Cham. https://doi.org/10.1007/978-3-030-47705-9_17

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  • DOI: https://doi.org/10.1007/978-3-030-47705-9_17

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