RGD-Functionalized Hydrogel Supports the Chondrogenic Commitment of Adipose Mesenchymal Stromal Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Experimental Plan
2.3. Raw Materials Characterization
2.3.1. FTIR Analysis
2.3.2. Hydrogel Preparation and Characterization
Rheological Characterization
Mechanical Characterization
Protein Diffusivity Study
2.4. hASCs Characterization
2.4.1. Cell Culture and Phenotypic Characterization of hASCs
2.4.2. hASCs-Laden Hydrogel Preparation and Biological Evaluations
2.4.3. Histology
2.4.4. Live/Dead Viability Assay
2.4.5. LDH Assay
2.4.6. Metabolic Activity Test
2.4.7. Chondrogenic Differentiation
2.4.8. Molecular Biology
2.4.9. Immunohistochemistry Staining
2.4.10. Statistical Analysis
3. Results and Discussion
3.1. Raw Materials Characterization: FTIR Analysis
3.2. Hydrogel Preparation and Characterization
3.2.1. Rheological Properties
3.2.2. Mechanical Properties
3.2.3. Protein Diffusivity Analysis
3.3. hASCs Characterization
3.3.1. Biological Evaluations of hASCs-Laden Hydrogels
3.3.2. RGD-Based Hydrogels Favor the Chondrogenic Processes
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Huey, D.J.; Hu, J.C.; Athanasiou, K.A. Unlike bone, cartilage regeneration remains elusive. Science 2012, 338, 917–921. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Cui, A.; Li, H.; Wang, D.; Zhong, J.; Chen, Y.; Lu, H. Global, regional prevalence, incidence and risk factors of knee osteoarthritis in population-based studies. EClinicalMedicine 2020, 29–30, 100587. [Google Scholar] [CrossRef] [PubMed]
- Krishnan, Y.; Grodzinsky, A.J. Cartilage diseases. Matrix Biol. 2018, 71–72, 51–69. [Google Scholar] [CrossRef] [PubMed]
- Kwon, H.; Brown, W.E.; Lee, C.A.; Wang, D.; Paschos, N.; Hu, J.C.; Athanasiou, K.A. Surgical and tissue engineering strategies for articular cartilage and meniscus repair. Nat. Rev. Rheumatol. 2019, 15, 550–570. [Google Scholar] [CrossRef]
- Izadifar, Z.; Chen, X.; Kulyk, W. Strategic design and fabrication of engineered scaffolds for articular cartilage repair. J. Funct. Biomater. 2012, 3, 799–838. [Google Scholar] [CrossRef] [Green Version]
- Armiento, A.R.; Stoddart, M.J.; Alini, M.; Eglin, D. Biomaterials for articular cartilage tissue engineering: Learning from biology. Acta Biomater. 2018, 65, 1–20. [Google Scholar] [CrossRef]
- Yang, J.; Zhang, Y.S.; Yue, K.; Khademhosseini, A. Cell-laden hydrogels for osteochondral and cartilage tissue engineering. Acta Biomater. 2017, 57, 1–25. [Google Scholar] [CrossRef]
- Bhattacharjee, M.; Coburn, J.; Centola, M.; Murab, S.; Barbero, A.; Kaplan, D.L.; Martin, I.; Ghosh, S. Tissue engineering strategies to study cartilage development, degeneration and regeneration. Adv. Drug Deliv. Rev. 2015, 84, 107–122. [Google Scholar] [CrossRef]
- Wei, W.; Ma, Y.; Yao, X.; Zhou, W.; Wang, X.; Li, C.; Lin, J.; He, Q.; Leptihn, S.; Ouyang, H. Advanced hydrogels for the repair of cartilage defects and regeneration. Bioact. Mater. 2021, 6, 998–1011. [Google Scholar] [CrossRef]
- Lin, H.; Yin, C.; Mo, A.; Hong, G. Applications of Hydrogel with Special Physical Properties in Bone and Cartilage Regeneration. Materials 2021, 14, 235. [Google Scholar] [CrossRef]
- Radhakrishnan, J.; Subramanian, A.; Krishnan, U.M.; Sethuraman, S. Injectable and 3D Bioprinted Polysaccharide Hydrogels: From Cartilage to Osteochondral Tissue Engineering. Biomacromolecules 2017, 18, 1–26. [Google Scholar] [CrossRef] [PubMed]
- Naghieh, S.; Sarker, M.; Sharma, N.K.; Barhoumi, Z.; Chen, X. Printability of 3D Printed Hydrogel Scaffolds: Influence of Hydrogel Composition and Printing Parameters. Appl. Sci. 2020, 10, 292. [Google Scholar] [CrossRef] [Green Version]
- Zhu, J.; Marchant, R.E. Design properties of hydrogel tissue-engineering scaffolds. Expert Rev. Med. Devices 2011, 8, 607–626. [Google Scholar] [CrossRef] [PubMed]
- Seidi, K.; Ayoubi-Joshaghani, M.H.; Azizi, M.; Javaheri, T.; Jaymand, M.; Alizadeh, E.; Webster, T.J.; Yazdi, A.A.; Niazi, M.; Hamblin, M.R.; et al. Bioinspired hydrogels build a bridge from bench to bedside. Nano Today 2021, 39, 101157. [Google Scholar] [CrossRef]
- Long, L.-Y.; Weng, Y.-X.; Wang, Y.-Z. Cellulose Aerogels: Synthesis, Applications, and Prospects. Polymers 2018, 10, 623. [Google Scholar] [CrossRef] [Green Version]
- Wei, Z.; Wu, C.; Li, R.; Yu, D.; Ding, Q. Nanocellulose based hydrogel or aerogel scaffolds for tissue engineering. Cellulose 2021, 28, 7497–7520. [Google Scholar] [CrossRef]
- Yahya, E.B.; Amirul, A.A.; Abdul Khalil, H.P.S.; Olaiya, N.G.; Iqbal, M.O.; Fauziah Jummaat, A.S.A.; Adnan, A.S. Insights into the Role of Biopolymer Aerogel Scaffolds in Tissue Engineering and Regenerative Medicine. Polymers 2021, 13, 1612. [Google Scholar] [CrossRef]
- Sánchez-Téllez, D.A.; Téllez-Jurado, L.; Rodríguez-Lorenzo, L.M. Hydrogels for Cartilage Regeneration, from Polysaccharides to Hybrids. Polymers 2017, 9, 671. [Google Scholar] [CrossRef] [Green Version]
- Nikolova, M.P.; Chavali, M.S. Recent advances in biomaterials for 3D scaffolds: A review. Bioact. Mater. 2019, 4, 271–292. [Google Scholar] [CrossRef]
- Zhao, T.; Wei, Z.; Zhu, W.; Weng, X. Recent Developments and Current Applications of Hydrogels in Osteoarthritis. Bioengineering 2022, 9, 132. [Google Scholar] [CrossRef]
- Lin, X.; Tsao, C.T.; Kyomoto, M.; Zhang, M. Injectable Natural Polymer Hydrogels for Treatment of Knee Osteoarthritis. Adv. Healthc. Mater. 2022, 11, e2101479. [Google Scholar] [CrossRef] [PubMed]
- Hafezi, M.; Nouri Khorasani, S.; Zare, M.; Esmaeely Neisiany, R.; Davoodi, P. Advanced Hydrogels for Cartilage Tissue Engineering: Recent Progress and Future Directions. Polymers 2021, 13, 4199. [Google Scholar] [CrossRef]
- Vega, S.L.; Kwon, M.; Mauck, R.L.; Burdick, J.A. Single Cell Imaging to Probe Mesenchymal Stem Cell N-Cadherin Mediated Signaling within Hydrogels. Ann. Biomed. Eng. 2016, 44, 1921–1930. [Google Scholar] [CrossRef] [Green Version]
- Foyt, D.A.; Taheem, D.K.; Ferreira, S.A.; Norman, M.D.A.; Petzold, J.; Jell, G.; Grigoriadis, A.E.; Gentleman, E. Hypoxia impacts human MSC response to substrate stiffness during chondrogenic differentiation. Acta Biomater. 2019, 89, 73–83. [Google Scholar] [CrossRef]
- Amann, E.; Wolff, P.; Breel, E.; van Griensven, M.; Balmayor, E.R. Hyaluronic acid facilitates chondrogenesis and matrix deposition of human adipose derived mesenchymal stem cells and human chondrocytes co-cultures. Acta Biomater. 2017, 52, 130–144. [Google Scholar] [CrossRef]
- Van Tomme, S.R.; Storm, G.; Hennink, W.E. In situ gelling hydrogels for pharmaceutical and biomedical applications. Int. J. Pharm. 2008, 355, 1–18. [Google Scholar] [CrossRef] [PubMed]
- Jeong, B.; Kim, S.W.; Bae, Y.H. Thermosensitive sol-gel reversible hydrogels. Adv. Drug Deliv. Rev. 2002, 54, 37–51. [Google Scholar] [CrossRef]
- Mandal, A.; Clegg, J.R.; Anselmo, A.C.; Mitragotri, S. Hydrogels in the clinic. Bioeng. Transl. Med. 2020, 5, e10158. [Google Scholar] [CrossRef] [Green Version]
- Huang, J.; Xiong, J.; Wang, D.; Zhang, J.; Yang, L.; Sun, S.; Liang, Y. 3D Bioprinting of Hydrogels for Cartilage Tissue Engineering. Gels 2021, 7, 144. [Google Scholar] [CrossRef] [PubMed]
- Yu, L.; Ding, J. Injectable hydrogels as unique biomedical materials. Chem. Soc. Rev. 2008, 37, 1473–1481. [Google Scholar] [CrossRef]
- Neves, S.C.; Moroni, L.; Barrias, C.C.; Granja, P.L. Leveling Up Hydrogels: Hybrid Systems in Tissue Engineering. Trends Biotechnol. 2020, 38, 292–315. [Google Scholar] [CrossRef]
- Boeuf, S.; Richter, W. Chondrogenesis of mesenchymal stem cells: Role of tissue source and inducing factors. Stem Cell Res. Ther. 2010, 1, 31. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Jorgensen, C.; Noël, D. Mesenchymal stem cells in osteoarticular diseases. Regen. Med. 2011, 6, 44–51. [Google Scholar] [CrossRef] [PubMed]
- Maumus, M.; Manferdini, C.; Toupet, K.; Peyrafitte, J.A.; Ferreira, R.; Facchini, A.; Gabusi, E.; Bourin, P.; Jorgensen, C.; Lisignoli, G.; et al. Adipose mesenchymal stem cells protect chondrocytes from degeneration associated with osteoarthritis. Stem Cell Res. 2013, 11, 834–844. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Manferdini, C.; Maumus, M.; Gabusi, E.; Piacentini, A.; Filardo, G.; Peyrafitte, J.A.; Jorgensen, C.; Bourin, P.; Fleury-Cappellesso, S.; Facchini, A.; et al. Adipose-derived mesenchymal stem cells exert antiinflammatory effects on chondrocytes and synoviocytes from osteoarthritis patients through prostaglandin E2. Arthritis Rheumatol. 2013, 65, 1271–1281. [Google Scholar] [CrossRef]
- Zoetebier, B.; Schmitz, T.; Ito, K.; Karperien, M.; Tryfonidou, M.A.; Paez, J. Injectable hydrogels for articular cartilage and nucleus pulposus repair: Status quo and prospects. Tissue Eng. Part A, 2022; ahead of print. [Google Scholar] [CrossRef]
- Yang, J.; Li, Y.; Liu, Y.; Li, D.; Zhang, L.; Wang, Q.; Xiao, Y.; Zhang, X. Influence of hydrogel network microstructures on mesenchymal stem cell chondrogenesis in vitro and in vivo. Acta Biomater. 2019, 91, 159–172. [Google Scholar] [CrossRef]
- Wang, F.; Nan, L.-P.; Zhou, S.-F.; Liu, Y.; Wang, Z.-Y.; Wang, J.-C.; Feng, X.-M.; Zhang, L. Injectable Hydrogel Combined with Nucleus Pulposus-Derived Mesenchymal Stem Cells for the Treatment of Degenerative Intervertebral Disc in Rats. Stem Cells Int. 2019, 2019, 8496025. [Google Scholar] [CrossRef]
- Li, Z.; Cao, B.; Wang, X.; Ye, K.; Li, S.; Ding, J. Effects of RGD nanospacing on chondrogenic differentiation of mesenchymal stem cells. J. Mater. Chem. B 2015, 3, 5197–5209. [Google Scholar] [CrossRef]
- Chen, F.; Ni, Y.; Liu, B.; Zhou, T.; Yu, C.; Su, Y.; Zhu, X.; Yu, X.; Zhou, Y. Self-crosslinking and injectable hyaluronic acid/RGD-functionalized pectin hydrogel for cartilage tissue engineering. Carbohydr. Polym. 2017, 166, 31–44. [Google Scholar] [CrossRef]
- Ruoslahti, E. RGD and other recognition sequences for integrins. Annu. Rev. Cell Dev. Biol. 1996, 12, 697–715. [Google Scholar] [CrossRef]
- Yang, M.; Zhang, Z.C.; Liu, Y.; Chen, Y.R.; Deng, R.H.; Zhang, Z.N.; Yu, J.K.; Yuan, F.Z. Function and Mechanism of RGD in Bone and Cartilage Tissue Engineering. Front. Bioeng. Biotechnol. 2021, 9, 773636. [Google Scholar] [CrossRef] [PubMed]
- Denier, J.P.; Dabrowski, P.P. On the boundary–layer equations for power–law fluids. Proc. R. Soc. Lond. Ser. A Math. Phys. Eng. Sci. 2004, 460, 3143–3158. [Google Scholar] [CrossRef]
- Blaeser, A.; Duarte Campos, D.F.; Puster, U.; Richtering, W.; Stevens, M.M.; Fischer, H. Controlling Shear Stress in 3D Bioprinting is a Key Factor to Balance Printing Resolution and Stem Cell Integrity. Adv. Healthc. Mater. 2016, 5, 326–333. [Google Scholar] [CrossRef] [PubMed]
- Trucco, D.; Vannozzi, L.; Teblum, E.; Telkhozhayeva, M.; Nessim, G.D.; Affatato, S.; Al-Haddad, H.; Lisignoli, G.; Ricotti, L. Graphene Oxide-Doped Gellan Gum–PEGDA Bilayered Hydrogel Mimicking the Mechanical and Lubrication Properties of Articular Cartilage. Adv. Healthc. Mater. 2021, 10, 2001434. [Google Scholar] [CrossRef]
- Manferdini, C.; Gabusi, E.; Sartore, L.; Dey, K.; Agnelli, S.; Almici, C.; Bianchetti, A.; Zini, N.; Russo, D.; Re, F.; et al. Chitosan-based scaffold counteracts hypertrophic and fibrotic markers in chondrogenic differentiated mesenchymal stromal cells. J. Tissue Eng. Regen. Med. 2019, 13, 1896–1911. [Google Scholar] [CrossRef]
- Dominici, M.; Le Blanc, K.; Mueller, I.; Slaper-Cortenbach, I.; Marini, F.; Krause, D.; Deans, R.; Keating, A.; Prockop, D.; Horwitz, E. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006, 8, 315–317. [Google Scholar] [CrossRef]
- Lisignoli, G.; Manferdini, C.; Lambertini, E.; Zini, N.; Angelozzi, M.; Gabusi, E.; Gambari, L.; Penolazzi, L.; Lolli, A.; Facchini, A.; et al. Chondrogenic potential of Slug-depleted human mesenchymal stem cells. Tissue Eng. Part A 2014, 20, 2795–2805. [Google Scholar] [CrossRef]
- Jin, M.; Shi, J.; Zhu, W.; Yao, H.; Wang, D.-A. Polysaccharide-Based Biomaterials in Tissue Engineering: A Review. Tissue Eng. Part B Rev. 2020, 27, 604–626. [Google Scholar] [CrossRef]
- O’Connell, G.; Garcia, J.; Amir, J. 3D Bioprinting: New Directions in Articular Cartilage Tissue Engineering. ACS Biomater. Sci. Eng. 2017, 3, 2657–2668. [Google Scholar] [CrossRef] [Green Version]
- Lee, H.-P.; Gu, L.; Mooney, D.J.; Levenston, M.E.; Chaudhuri, O. Mechanical confinement regulates cartilage matrix formation by chondrocytes. Nat. Mater. 2017, 16, 1243–1251. [Google Scholar] [CrossRef] [Green Version]
- Cherne, M.D.; Sidar, B.; Sebrell, T.A.; Sanchez, H.S.; Heaton, K.; Kassama, F.J.; Roe, M.M.; Gentry, A.B.; Chang, C.B.; Walk, S.T.; et al. A Synthetic Hydrogel, VitroGel® ORGANOID-3, Improves Immune Cell-Epithelial Interactions in a Tissue Chip Co-Culture Model of Human Gastric Organoids and Dendritic Cells. Front. Pharmacol. 2021, 12, 707891. [Google Scholar] [CrossRef] [PubMed]
- Baltazar, T.; Jiang, B.; Moncayo, A.; Merola, J.; Albanna, M.Z.; Saltzman, W.M.; Pober, J.S. 3D bioprinting of an implantable xeno-free vascularized human skin graft. Bioeng. Transl. Med. 2022, e10324. [Google Scholar] [CrossRef]
- Firner, S.; Zaucke, F.; Michael, J.; Dargel, J.; Schiwy-Bochat, K.H.; Heilig, J.; Rothschild, M.A.; Eysel, P.; Brüggemann, G.P.; Niehoff, A. Extracellular Distribution of Collagen II and Perifibrillar Adapter Proteins in Healthy and Osteoarthritic Human Knee Joint Cartilage. J. Histochem. Cytochem. 2017, 65, 593–606. [Google Scholar] [CrossRef] [PubMed]
- Heinegård, D. Fell-Muir Lecture: Proteoglycans and more—From molecules to biology. Int. J. Exp. Pathol. 2009, 90, 575–586. [Google Scholar] [CrossRef]
- Chen, F.H.; Herndon, M.E.; Patel, N.; Hecht, J.T.; Tuan, R.S.; Lawler, J. Interaction of cartilage oligomeric matrix protein/thrombospondin 5 with aggrecan. J. Biol. Chem. 2007, 282, 24591–24598. [Google Scholar] [CrossRef] [Green Version]
- Liu, S.Q.; Tian, Q.; Wang, L.; Hedrick, J.L.; Hui, J.H.P.; Yang, Y.Y.; Ee, P.L.R. Injectable Biodegradable Poly(ethylene glycol)/RGD Peptide Hybrid Hydrogels for in vitro Chondrogenesis of Human Mesenchymal Stem Cells. Macromol. Rapid Commun. 2010, 31, 1148–1154. [Google Scholar] [CrossRef]
- Li, S.; Wang, X.; Cao, B.; Ye, K.; Li, Z.; Ding, J. Effects of Nanoscale Spatial Arrangement of Arginine-Glycine-Aspartate Peptides on Dedifferentiation of Chondrocytes. Nano Lett. 2015, 15, 7755–7765. [Google Scholar] [CrossRef]
- Villanueva, I.; Weigel, C.; Bryant, S. Cell–matrix interactions and dynamic mechanical loading influence chondrocyte gene expression and bioactivity in PEG-RGD hydrogels. Acta Biomater. 2009, 5, 2832–2846. [Google Scholar] [CrossRef] [Green Version]
- Salinas, C.N.; Anseth, K.S. The enhancement of chondrogenic differentiation of human mesenchymal stem cells by enzymatically regulated RGD functionalities. Biomaterials 2008, 29, 2370–2377. [Google Scholar] [CrossRef] [Green Version]
- Kloxin, A.M.; Kasko, A.M.; Salinas, C.N.; Anseth, K.S. Photodegradable hydrogels for dynamic tuning of physical and chemical properties. Science 2009, 324, 59–63. [Google Scholar] [CrossRef] [Green Version]
- Di Maggio, N.; Martella, E.; Frismantiene, A.; Resink, T.J.; Schreiner, S.; Lucarelli, E.; Jaquiery, C.; Schaefer, D.J.; Martin, I.; Scherberich, A. Extracellular matrix and α(5)β(1) integrin signaling control the maintenance of bone formation capacity by human adipose-derived stromal cells. Sci. Rep. 2017, 7, 44398. [Google Scholar] [CrossRef] [PubMed] [Green Version]
VG-3D 1:1 | VG-3D 1:2 | VG-RGD 1:1 | VG-RGD 1:2 | |
---|---|---|---|---|
K [Pa·sn] | 3.36 ± 0.69 | 1.56 ± 0.52 | 2.03 ± 0.38 | 1.12 ± 0.36 |
n | 0.21 ± 0.02 | 0.22 ± 0.09 | 0.20 ± 0.03 | 0.29 ± 0.04 |
τ [Pa] | 27.12 ± 9.01 | 15.38 ± 8.24 | 21.33 ± 8.48 | 15.93 ± 5.12 |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Manferdini, C.; Trucco, D.; Saleh, Y.; Gabusi, E.; Dolzani, P.; Lenzi, E.; Vannozzi, L.; Ricotti, L.; Lisignoli, G. RGD-Functionalized Hydrogel Supports the Chondrogenic Commitment of Adipose Mesenchymal Stromal Cells. Gels 2022, 8, 382. https://doi.org/10.3390/gels8060382
Manferdini C, Trucco D, Saleh Y, Gabusi E, Dolzani P, Lenzi E, Vannozzi L, Ricotti L, Lisignoli G. RGD-Functionalized Hydrogel Supports the Chondrogenic Commitment of Adipose Mesenchymal Stromal Cells. Gels. 2022; 8(6):382. https://doi.org/10.3390/gels8060382
Chicago/Turabian StyleManferdini, Cristina, Diego Trucco, Yasmin Saleh, Elena Gabusi, Paolo Dolzani, Enrico Lenzi, Lorenzo Vannozzi, Leonardo Ricotti, and Gina Lisignoli. 2022. "RGD-Functionalized Hydrogel Supports the Chondrogenic Commitment of Adipose Mesenchymal Stromal Cells" Gels 8, no. 6: 382. https://doi.org/10.3390/gels8060382