Home A versatile perfusion bioreactor and endothelializable photo cross-linked tubes of gelatin methacryloyl as promising tools in tissue engineering
Article
Licensed
Unlicensed Requires Authentication

A versatile perfusion bioreactor and endothelializable photo cross-linked tubes of gelatin methacryloyl as promising tools in tissue engineering

  • Birgit Huber , Eva Hoch , Iván Calderon , Kirsten Borchers and Petra J. Kluger EMAIL logo
Published/Copyright: September 18, 2018

Abstract

Size and function of bioartificial tissue models are still limited due to the lack of blood vessels and dynamic perfusion for nutrient supply. In this study, we evaluated the use of cytocompatible methacryl-modified gelatin for the fabrication of a hydrogel-based tube by dip-coating and subsequent photo-initiated cross-linking. The wall thickness of the tubes and the diameter were tuned by the degree of gelatin methacryl-modification and the number of dipping cycles. The dipping temperature of the gelatin solution was adjusted to achieve low viscous fluids of approximately 0.1 Pa s and was different for gelatin derivatives with different modification degrees. A versatile perfusion bioreactor for the supply of surrounding tissue models was developed, which can be adapted to several geometries and sizes of blood-vessel mimicking tubes. The manufactured bendable gelatin tubes were permeable for water and dissolved substances, like Nile Blue and serum albumin. As a proof of concept, human fibroblasts in a three-dimensional collagen tissue model were successfully supplied with nutrients via the central gelatin tube under dynamic conditions for 2 days. Moreover, the tubes could be used as scaffolds to build-up a functional and viable endothelial layer. Hence, the presented tools can contribute to solving current challenges in tissue engineering.

Acknowledgments

The authors thank Sarah Schmidt and Regina Buck for the preparation of the gelatin hydrogel tubes. Warm acknowledgments also go to Eva Conraths, Kirstin Linke and Antonia Link for their extraordinary helpful support in the labs.

  1. Author Statement

  2. Research funding: This work was supported by the European Commission under the Seventh Framework Program, Funder Id: 10.13039/100011102 (grant agreement no. 263416).

  3. Conflict of interest: Authors state no conflict of interest.

  4. Informed consent: All patients gave a written agreement according to the permission of the Landesärztekammer Baden-Württemberg (F-2012-078; for normal skin from elective surgeries).

  5. Ethical approval: The research related to human use complied with all the relevant national regulations and institutional policies, was performed in accordance with the tenets of the Helsinki Declaration, and has been approved by the local Ethics Committee.

References

[1] Khademhosseini A, Langer R, Borenstein J, Vacanti JP. Microscale technologies for tissue engineering and biology. Proc Natl Acad Sci USA 2006;103:2480–7.10.1073/pnas.0507681102Search in Google Scholar PubMed PubMed Central

[2] Langer R, Vacanti JP. Tissue engineering. Science 1993;260:920–6.10.1126/science.8493529Search in Google Scholar PubMed

[3] Carmeliet P, Jain RK. Angiogenesis in cancer and other diseases. Nature 2000;407:249–57.10.1038/35025220Search in Google Scholar PubMed

[4] Kannan RY, Salacinski HJ, Sales K, Butler P, Seifalian AM. The roles of tissue engineering and vascularisation in the development of micro-vascular networks: a review. Biomaterials 2005;26:1857–75.10.1016/j.biomaterials.2004.07.006Search in Google Scholar PubMed

[5] Novosel EC, Kleinhans C, Kluger PJ. Vascularization is the key challenge in tissue engineering. Adv Drug Deliv Rev 2011;63:300–11.10.1016/j.addr.2011.03.004Search in Google Scholar PubMed

[6] Burton AC. Relation of structure to function of the tissues of the wall of blood vessels. Physiol Rev 1954;34:619–42.10.1152/physrev.1954.34.4.619Search in Google Scholar PubMed

[7] Syazwani N, Azhim A, Morimoto Y, Furukawa KS, Ushida T. Decellularization of aorta tissue using sonication treatment as potential scaffold for vascular tissue engineering. J Med Biol Eng 2015;35:258–69.10.1007/s40846-015-0028-5Search in Google Scholar

[8] L’Heureux N, Pâquet S, Labbé R, Germain L, Auger FA. A completely biological tissue-engineered human blood vessel. FASEB J 1998;12:47–56.10.1096/fasebj.12.1.47Search in Google Scholar PubMed

[9] Hoch E, Tovar GE, Borchers K. Bioprinting of artificial blood vessels: current approaches towards a demanding goal. Eur J Cardiothorac Surg 2014;46:767–78.10.1093/ejcts/ezu242Search in Google Scholar PubMed

[10] Datta P, Ayan B, Ozbolat IT. Bioprinting for vascular and vascularized tissue biofabrication. Acta Biomater 2017;51:1–20.10.1016/j.actbio.2017.01.035Search in Google Scholar PubMed

[11] Huber B, Engelhardt S, Meyer W, Krüger H, Wenz A, Schönhaar V, et al. Blood-vessel mimicking structures by stereolithographic fabrication of small porous tubes using cytocompatible polyacrylate elastomers, biofunctionalization and endothelialization. J Funct Biomater 2016;7:11.10.3390/jfb7020011Search in Google Scholar PubMed PubMed Central

[12] Baudis S, Nehl F, Ligon SC, Nigisch A, Bergmeister H, Bernhard D, et al. Elastomeric degradable biomaterials by photopolymerization-based CAD-CAM for vascular tissue engineering. Biomed Mater 2011;6:055003.10.1088/1748-6041/6/5/055003Search in Google Scholar PubMed

[13] Nguyen DH, Stapleton SC, Yang MT, Cha SS, Choi CK, Galie PA, et al. Biomimetic model to reconstitute angiogenic sprouting morphogenesis in vitro. Proc Natl Acad Sci USA 2013;110:6712–7.10.1073/pnas.1221526110Search in Google Scholar PubMed PubMed Central

[14] Asakawa N, Shimizu T, Tsuda Y, Sekiya S, Sasagawa T, Yamato M, et al. Pre-vascularization of in vitro three-dimensional tissues created by cell sheet engineering. Biomaterials 2010;31:3903–9.10.1016/j.biomaterials.2010.01.105Search in Google Scholar PubMed

[15] Miller JS, Stevens KR, Yang MT, Baker BM, Nguyen DH, Cohen DM, et al. Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues. Nat Mater 2012;11:768–74.10.1038/nmat3357Search in Google Scholar PubMed PubMed Central

[16] Sekine H, Shimizu T, Sakaguchi K, Dobashi I, Wada M, Yamato M, et al. In vitro fabrication of functional three-dimensional tissues with perfusable blood vessels. Nat Commun 2013;4:1399.10.1038/ncomms2406Search in Google Scholar PubMed PubMed Central

[17] Tocchio A, Tamplenizza M, Martello F, Gerges I, Rossi E, Argentiere S, et al. Versatile fabrication of vascularizable scaffolds for large tissue engineering in bioreactor. Biomaterials 2015;45:124–31.10.1016/j.biomaterials.2014.12.031Search in Google Scholar PubMed

[18] Sakaguchi K, Shimizu T, Horaguchi S, Sekine H, Yamato M, Umezu M, et al. In vitro engineering of vascularized tissue surrogates. Sci Rep 2013;3:1316.10.1038/srep01316Search in Google Scholar PubMed PubMed Central

[19] Hoch E, Schuh C, Hirth T, Tovar GE, Borchers K. Stiff gelatin hydrogels can be photo-chemically synthesized from low viscous gelatin solutions using molecularly functionalized gelatin with a high degree of methacrylation. J Mater Sci Mater Med 2012;23:2607–17.10.1007/s10856-012-4731-2Search in Google Scholar PubMed

[20] Hoch E, Hirth T, Tovar GE, Borchers K. Chemical tailoring of gelatin to adjust its chemical and physical properties for functional bioprinting. J Mater Chem B 2013;1:5675–85.10.1039/c3tb20745eSearch in Google Scholar PubMed

[21] Wenz A, Janke K, Hoch E, Tovar GE, Borchers K, Kluger PJ. Hydroxyapatite-modified gelatin bioinks for bone bioprinting. BioNanoMaterials 2016;17:179.10.1515/bnm-2015-0018Search in Google Scholar

[22] Wenz A, Borchers K, Tovar GE, Kluger PJ. Bone matrix production in hydroxyapatite-modified hydrogels suitable for bone bioprinting. Biofabrication 2017;9:044103.10.1088/1758-5090/aa91ecSearch in Google Scholar PubMed

[23] Claaßen C, Claaßen MH, Truffault V, Sewald L, Tovar GE, Borchers K, et al. Quantification of substitution of gelatin methacryloyl: best practice and current pitfalls. Biomacromolecules 2018;19:42–52.10.1021/acs.biomac.7b01221Search in Google Scholar PubMed

[24] Schuerlein S, Schwarz T, Krziminski S, Gätzner S, Hoppensack A, Schwedhelm I, et al. A versatile modular bioreactor platform for tissue engineering. Biotech J 2017;12:1600326.10.1002/biot.201600326Search in Google Scholar PubMed PubMed Central

[25] Stephenson M, Grayson W. Recent advances in bioreactors for cell-based therapies. F1000Res 2018;7:517.10.12688/f1000research.12533.1Search in Google Scholar PubMed PubMed Central

[26] Wolf F, Rojas González DM, Steinseifer U, Obdenbusch M, Herfs W, Brecher C, et al. VascuTrainer: a mobile and disposable bioreactor system for the conditioning of tissue-engineered vascular grafts. Ann Biomed Eng 2018;46:616–26.10.1007/s10439-018-1977-ySearch in Google Scholar PubMed

[27] Van Den Bulcke AI, Bogdanov B, De Rooze N, Schacht EH, Cornelissen M, Berghmans H. Structural and rheological properties of methacrylamide modified gelatin hydrogels. Biomacromolecules 2000;1:31–8.10.1021/bm990017dSearch in Google Scholar PubMed

[28] Fairbanks BD, Schwartz MP, Bowman CN, Anseth KS. Photoinitiated polymerization of PEG-diacrylate with lithium phenyl-2,4,6-trimethylbenzoylphosphinate: polymerization rate and cytocompatibility. Biomaterials 2009;30:6702–7.10.1016/j.biomaterials.2009.08.055Search in Google Scholar PubMed PubMed Central

[29] Engelhardt S, Hoch E, Borchers K, Meyer W, Krüger H, Tovar G, et al. Fabrication of 2D protein microstructures and 3D polymer–protein hybrid microstructures by two-photon polymerization. Biofabrication 2011;3:025003.10.1088/1758-5082/3/2/025003Search in Google Scholar PubMed

[30] Kluger PJ, Wyrwa R, Weisser J, Maierle J, Votteler M, Rode C, et al. Electrospun poly(D/L-lactide-co-L-lactide) hybrid matrix: a novel scaffold material for soft tissue engineering. J Mater Sci Mater Med 2010;21:2665–71.10.1007/s10856-010-4128-zSearch in Google Scholar PubMed PubMed Central

[31] Huber B, Czaja AM, Kluger PJ. Influence of epidermal growth factor (EGF) and hydrocortisone on the co-culture of mature adipocytes and endothelial cells for vascularized adipose tissue engineering. Cell Biol Int 2016;40:569–78.10.1002/cbin.10595Search in Google Scholar PubMed

[32] Schrieber R, Gareis H. Gelatine Handbook. Weinheim, Germany: WILEY-VCH Verlag GmbH & Co.; 2007.10.1002/9783527610969Search in Google Scholar

[33] Huber B, Link A, Linke K, Gehrke SA, Winnefeld M, Kluger PJ. Integration of mature adipocytes to build-up a functional three-layered full-skin equivalent. Tissue Eng Part C Methods 2016;22:756–64.10.1089/ten.tec.2016.0141Search in Google Scholar PubMed PubMed Central

[34] Hahn MS, McHale MK, Wang E, Schmedlen RH, West JL. Physiologic pulsatile flow bioreactor conditioning of poly(ethylene glycol)-based tissue engineered vascular grafts. Ann Biomed Eng 2007;35:190–200.10.1007/s10439-006-9099-3Search in Google Scholar PubMed

[35] Iwasaki K, Kojima K, Kodama S, Paz AC, Chambers M, Umezu M, et al. Bioengineered three-layered robust and elastic artery using hemodynamically-equivalent pulsatile bioreactor. Circulation 2008;118:S52–7.10.1161/CIRCULATIONAHA.107.757369Search in Google Scholar PubMed

[36] Cardinal KO, Bonnema GT, Hofer H, Barton JK, Williams SK. Tissue-engineered vascular grafts as in vitro blood vessel mimics for the evaluation of endothelialization of intravascular devices. Tissue Eng 2006;12:3431–8.10.1089/ten.2006.12.3431Search in Google Scholar PubMed

[37] Xu J, Ge H, Zhou X, Yang D, Guo T, He J, et al. Tissue-engineered vessel strengthens quickly under physiological deformation: application of a new perfusion bioreactor with machine vision. J Vasc Res 2005;42:503–8.10.1159/000088161Search in Google Scholar PubMed

[38] Groeber F, Kahlig A, Loff S, Walles H, Hansmann J. A bioreactor system for interfacial culture and physiological perfusion of vascularized tissue equivalents. Biotechnol J 2013;8:308–16.10.1002/biot.201200160Search in Google Scholar PubMed

[39] Ward A, Quinn KP, Bellas E, Georgakoudi I, Kaplan DL. Noninvasive metabolic imaging of engineered 3D human adipose tissue in a perfusion bioreactor. PLoS One 2013;8:e55696.10.1371/journal.pone.0055696Search in Google Scholar PubMed PubMed Central

[40] Villalona GA, Udelsman B, Duncan DR, McGillicuddy E, Sawh-Martinez RF, Hibino N, et al. Cell-seeding techniques in vascular tissue engineering. Tissue Eng Part B Rev 2010;16:341–50.10.1089/ten.teb.2009.0527Search in Google Scholar PubMed PubMed Central

[41] Soletti L, Nieponice A, Guan J, Stankus JJ, Wagner WR, Vorp DA. A seeding device for tissue engineered tubular structures. Biomaterials 2006;27:4863–70.10.1016/j.biomaterials.2006.04.042Search in Google Scholar PubMed

Received: 2018-01-29
Accepted: 2018-08-06
Published Online: 2018-09-18
Published in Print: 2019-08-27

©2019 Walter de Gruyter GmbH, Berlin/Boston

Downloaded on 12.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/bmt-2018-0015/html
Scroll to top button