Extracellular matrix (ECM) has a major role in the structural support and cellular processes of organs and tissues. Proteins extracted from the ECM have been used to fabricate different scaffolds for tissue engineering applications. The aims of the present study were to extract, characterize and fabricate a new class of hydrogel with proteins isolated from pig bone ECM and combine them with a synthetic polymer so it could be used to promote bone regeneration. Porcine bone demineralized and digested extracellular matrix (pddECM) containing collagen type I was produced, optimized and sterilized with high pressurized CO2 method. The pddECM was further blended with 20% w/v polyethylene glycol diacrylate (PEGDA) to create an injectable semi interpenetrating polymer network (SIPN) scaffold with enhanced physicochemical properties. The blend tackled the shortfall of natural polymers, such as lack of structural stability and fast degradation, preserving its structure in more than 90% after 30 days of incubation; thus, increasing the material endurance in a simulated physiological environment. The manufactured injectable hydrogel showed high cytocompatibility with hOb and SaOs-2 cells, promoting osteogenic proliferation within 21 days of culture. The hydrogel had a high compression modulus of 520 kPa, low swelling (5.3 mg/mg) and millimetric volume expansion (19.5%), all of which are favorable characteristics for bone regeneration applications.
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Bonafede M, Espindle D, Bower AG. The direct and indirect costs of long bone fractures in a working age US population. J Med Econ. 2013;16:169–78.
Delea TE, McKiernan J, Brandman J, Edelsberg J, Sung J, Raut M, et al. Impact of skeletal complications on total medical care costs among patients with bone metastases of lung cancer. J Thorac Oncol. 2006;1:571–6.
Woolf AD, Pfleger B. Burden of major musculoskeletal conditions. Bull World Health Organ. 2003;81:646–56.
Deev RV, Drobyshev AY, Bozo IY, Isaev AA. Ordinary and activated bone grafts: applied classification and the main features. BioMed Res Int. 2015;2015:365050.
Nauth A, Lane J, Watson JT, Giannoudis P. Bone graft substitution and augmentation. J Orthop Trauma. 2015;29:S34–8.
Egol KA, Nauth A, Lee M, Pape HC, Watson JT, Borrelli J Jr. Bone grafting: sourcing, timing, strategies, and alternatives. J Orthop Trauma. 2015;29:S10–4.
Van Lieshout EM, Alt V. Bone graft substitutes and bone morphogenetic proteins for osteoporotic fractures: what is the evidence. Injury. 2016;47:S43–6.
Delloye C, Cornu O, Druez V, Barbier O. Bone allografts: what they can offer and what they cannot. J Bone Jt Surg Br. 2007;89:574–9.
Graham SM, Leonidou A, Aslam-Pervez N, Hamza A, Panteliadis P, Heliotis M, et al. Biological therapy of bone defects: the immunology of bone allo-transplantation. Expert Opin Biol Ther. 2010;10:885–901.
Amini AR, Laurencin CT, Nukavarapu SP. Bone tissue engineering: recent advances and challenges. Crit Rev Biomed Eng. 2012;40:363–408.
Ullah F, Othman MB, Javed F, Ahmad Z, Md Akil H. Classification, processing and application of hydrogels: a review. Mater Sci Eng C, Mater Biol Appl. 2015;57:414–33.
Badylak SF. The extracellular matrix as a biologic scaffold material. Biomaterials.2007;28:3587–93.
Boden SD, Schimandle JH, Hutton WC. Lumbar intertransverse-process spinal arthrodesis with use of a bovine bone-derived osteoinductive protein. A preliminary report. J bone Jt Surg Am. 1995;77:1404–17.
Geckil H, Xu F, Zhang X, Moon S, Demirci U. Engineering hydrogels as extracellular matrix mimics. Nanomedicine. 2010;5:469–84.
Fitzpatrick LE, McDevitt TC. Cell-derived matrices for tissue engineering and regenerative medicine applications. Biomater Sci. 2015;3:12–24.
Rosso F, Giordano A, Barbarisi M, Barbarisi A. From cell-ECM interactions to tissue engineering. J Cell Physiol. 2004;199:174–80.
Kular JK, Basu S, Sharma RI. The extracellular matrix: structure, composition, age-related differences, tools for analysis and applications for tissue engineering. J Tissue Eng. 2014;5:2041731414557112.
Freytes DO, Martin J, Velankar SS, Lee AS, Badylak SF. Preparation and rheological characterization of a gel form of the porcine urinary bladder matrix. Biomaterials. 2008;29:1630–7.
Greco KV, Francis L, Somasundaram M, Greco G, English NR, Roether JA, et al. Characterisation of porcine dermis scaffolds decellularised using a novel non-enzymatic method for biomedical applications. J Biomater Appl. 2015;30:239–53.
Sawkins MJ, Bowen W, Dhadda P, Markides H, Sidney LE, Taylor AJ, et al. Hydrogels derived from demineralized and decellularized bone extracellular matrix. Acta Biomater. 2013;9:7865–73.
Williams C, Budina E, Stoppel WL, Sullivan KE, Emani S, Emani SM, et al. Cardiac extracellular matrix-fibrin hybrid scaffolds with tunable properties for cardiovascular tissue engineering. Acta Biomater. 2015;14:84–95.
Choi B, Kim S, Lin B, Wu BM, Lee M. Cartilaginous extracellular matrix-modified chitosan hydrogels for cartilage tissue engineering. ACS Appl Mater interfaces. 2014;6:20110–21.
Mehta M, Madl CM, Lee S, Duda GN, Mooney DJ. The collagen I mimetic peptide DGEA enhances an osteogenic phenotype in mesenchymal stem cells when presented from cell-encapsulating hydrogels. J Biomed Mater Res Part A. 2015;103:3516–25.
Yang M, Wang J, Zhu Y, Mao C. Bio-templated growth of bone minerals from modified simulated body fluid on nanofibrous decellularized natural tissues. J Biomed Nanotechnol. 2016;12:753–61.
Ramshaw JA, Peng YY, Glattauer V, Werkmeister JA. Collagens as biomaterials. J Mater Sci Mater Med. 2009;20:S3–8.
Chevallay B, Herbage D. Collagen-based biomaterials as 3D scaffold for cell cultures: applications for tissue engineering and gene therapy. Med Biol Eng Comput. 2000;38:211–8.
McNaught AD, Wilkinson A. IUPAC. Compendium of chemical terminology, 2nd revised ed. (The “Gold Book”). Cambridge, UK: Wiley-Blackwell scientific publications, Royal Society of Chemistry; 1997.
Zhang JT, Bhat R, Jandt KD. Temperature-sensitive PVA/PNIPAAm semi-IPN hydrogels with enhanced responsive properties. Acta biomaterialia. 2009;5:488–97.
Fathi A, Lee S, Breen A, Shirazi AN, Valtchev P, Dehghani F. Enhancing the mechanical properties and physical stability of biomimetic polymer hydrogels for micro-patterning and tissue engineering applications. Eur Polym J. 2014;59:161–70.
Fathi A, Lee S, Zhong X, Hon N, Valtchev P, Dehghani F. Fabrication of interpenetrating polymer network to enhance the biological activity of synthetic hydrogels. Polymer. 2013;54:5534–42.
Hong Y, Huber A, Takanari K, Amoroso NJ, Hashizume R, Badylak SF, et al. Mechanical properties and in vivo behavior of a biodegradable synthetic polymer microfiber-extracellular matrix hydrogel biohybrid scaffold. Biomaterials. 2011;32:3387–94.
Martin I, Garcia T, Fajardo V, Rojas M, Pegels N, Hernandez PE, et al. SYBR-Green real-time PCR approach for the detection and quantification of pig DNA in feedstuffs. Meat Sci. 2009;82:252–9.
Dillow AK, Dehghani F, Hrkach JS, Foster NR, Langer R. Bacterial inactivation by using near- and supercritical carbon dioxide. Proc Natl Acad Sci USA. 1999;96:10344–8.
Torabian G, Bahramian B, Zambon A, Spilimbergo S, Adil Q, Schindeler A, et al. A hybrid process for increasing the shelf life of elderberry juice. J Supercrit Fluids. 2018;140:406–14.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680–5.
Rathsam C, Eaton RE, Simpson CL, Browne GV, Berg T, Harty DW, et al. Up-regulation of competence- but not stress-responsive proteins accompanies an altered metabolic phenotype in Streptococcus mutans biofilms. Microbiology. 2005;151:1823–37.
Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes. Biomaterials. 2011;32:3233–43.
Black CR, Goriainov V, Gibbs D, Kanczler J, Tare RS, Oreffo RO. Bone tissue engineering. Curr Mol Biol Rep. 2015;1:132–40.
Miller EJ, Rhodes RK. Preparation and characterization of the different types of collagen. Methods Enzymol. 1982;82:33–64.
Pietrzak WS, Ali SN, Chitturi D, Jacob M, Woodell-May JE. BMP depletion occurs during prolonged acid demineralization of bone: characterization and implications for graft preparation. Cell Tissue Bank. 2011;12:81–8.
Delgado LM, Pandit A, Zeugolis DI. Influence of sterilisation methods on collagen-based devices stability and properties. Expert Rev Med Devices. 2014;11:305–14.
Russell AD, Furr JR, Maillard JY. Synergistic sterilization. PDA J Pharm Sci Technol. 1997;51:174–5.
Mellonig JT, Prewett AB, Moyer MP. HIV inactivation in a bone allograft. J Periodontol. 1992;63:979–83.
Ehrbar M, Sala A, Lienemann P, Ranga A, Mosiewicz K, Bittermann A, et al. Elucidating the role of matrix stiffness in 3D cell migration and remodeling. Biophys J. 2011;100:284–93.
Bryant SJ, Bender RJ, Durand KL, Anseth KS. Encapsulating chondrocytes in degrading PEG hydrogels with high modulus: engineering gel structural changes to facilitate cartilaginous tissue production. Biotechnol Bioeng. 2004;86:747–55.
Nair LS, Laurencin CT. Biodegradable polymers as biomaterials. Prog Polym Sci. 2007;32:762–98.
Yu T, Wang W, Nassiri S, Kwan T, Dang C, Liu W, et al. Temporal and spatial distribution of macrophage phenotype markers in the foreign body response to glutaraldehyde-crosslinked gelatin hydrogels. J Biomater Sci Polym Ed. 2016;27:721–42.
Khademhosseini A, Langer R. Microengineered hydrogels for tissue engineering. Biomaterials. 2007;28:5087–92.
da Luz Moreira P, Genari SC, Goissis G, Galembeck F, An YH, Santos AR Jr. Bovine osteoblasts cultured on polyanionic collagen scaffolds: an ultrastructural and immunocytochemical study. J Biomed Mater Res Part B, Appl Biomater. 2013;101:18–27.
The authors would like to acknowledge the support from, The University of Sydney Scool of Dentistry and Chemical and Biomolecular Engineering, The University of Sydney, Mass Spectrometry Facility Core and Scanning Electron Microscopy facility, Sydney, NSW, Australia, Australian Dental Research Foundation (ADRF) 40-2015 CONICYT PAI/INDUSTRIA 79090016, 3M ESPE, and the technical support of Elizabeth Kelly, Filip Vujovic and Mara Cvejic.
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Obregon-Miano, F., Fathi, A., Rathsam, C. et al. Injectable porcine bone demineralized and digested extracellular matrix—PEGDA hydrogel blend for bone regeneration. J Mater Sci: Mater Med 31, 21 (2020). https://doi.org/10.1007/s10856-019-6354-3