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Strategies on process engineering of chondrocyte culture for cartilage tissue regeneration

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Abstract

The current work is an attempt to study the strategies for cartilage tissue regeneration using porous scaffold in wavy walled airlift bioreactor (ALBR). Novel chitosan, poly (l-lactide) and hyaluronic acid based composite scaffold were prepared. The scaffolds were cross-linked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, N-hydroxysuccinimide and chondroitin sulfate to obtain interconnected 3D microstructure showing excellent biocompatibility, higher cellular differentiation and increased stability. The surface morphology and porosity of the scaffolds were analyzed using scanning electron microscopy (SEM) and mercury intrusion porosimeter and optimized for chondrocyte regeneration. The study shows that the scaffolds were highly porous with pore size ranging from 48 to 180 µm and the porosities in the range 80–92%. Swelling and in vitro degradation studies were performed for the composite scaffolds; by increasing the chitosan: HA ratio in the composite scaffolds, the swelling property increases and stabilizes after 24 h. There was controlled degradation of composite scaffolds for 4 weeks. The uniform chondrocyte distribution in the scaffold using various growth modes in the shake flask and ALBR was studied by glycosaminoglycans (GAG) quantification, MTT assay and mixing time evaluation. The cell culture studies demonstrated that efficient designing of ALBR increases the cartilage regeneration as compared to using a shake flask. The free chondrocyte microscopy and cell attachment were performed by inverted microscope and SEM, and from the study it was confirmed that the cells uniformly attached to the scaffold. This study focuses on optimizing strategies for the culture of chondrocyte using suitable scaffold for improved cartilage tissue regeneration.

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References

  1. Griffith LG, Naughton G (2002) Tissue engineering—current challenges and expanding opportunities. Science 295:1009–1014

    Article  CAS  Google Scholar 

  2. Guilak F, Butler DL, Goldstein SA (2001) Functional tissue engineering: the role of biomechanics in articular cartilage repair. Clin Orthop Relat Res 391:S295–S305

    Article  Google Scholar 

  3. Kaur T, Thirugnanam A (2016) Tailoring in vitro biological and mechanical properties of polyvinyl alcohol reinforced with threshold carbon nanotube concentration for improved cellular response. RSC Advances 6:39982–39992

    Article  CAS  Google Scholar 

  4. Maiti P, Patel D, Singh RK, Singh S, Aswal VK, Rana D, Ray B (2016) Graphene as chain extender of polyurethanes for biomedical applications. RSC Adv 6:58628–58640

    Article  Google Scholar 

  5. Naseri N, Poirier J-M, Girandon L, Fröhlich M, Oksman K, Mathew AP (2016) 3-Dimensional porous nanocomposite scaffolds based on cellulose nanofibers for cartilage tissue engineering: tailoring of porosity and mechanical performance. RSC Adv 6:5999–6007

    Article  CAS  Google Scholar 

  6. Pathria MN, Chung CB, Resnick DL (2016) Acute and stress-related injuries of bone and cartilage: pertinent anatomy, basic biomechanics, and imaging perspective. Radiology 280:21–38

    Article  Google Scholar 

  7. Sharma C, Gautam S, Dinda AK, Mishra NC (2011) Cartilage tissue engineering: current scenario and challenges. Adv Mater Lett 2:90–99

    Article  CAS  Google Scholar 

  8. Tibbitt MW, Rodell CB, Burdick JA, Anseth KS (2015) Progress in material design for biomedical applications. Proc Natl Acad Sci 112:14444–14451

    Article  CAS  Google Scholar 

  9. Schussler S, Guiro K, Livingston T (2015) In: Antoniac IV (ed) Handbook of bioceramics and biocomposites. Springer, Switzerland

  10. Nandgaonkar A, Krause W, Lucia L (2016) Nanocomposite for musculoskeletal tissue regeneration. Woodhead publishing series in biomaterials

  11. Ching KY, Andriotis OG, Li S, Basnett P, Su B, Roy I, Tare RS, Sengers BG, Stolz M (2016) Nanofibrous poly (3-hydroxybutyrate)/poly (3-hydroxyoctanoate) scaffolds provide a functional microenvironment for cartilage repair. J Biomater Appl 31:77–91

    Article  CAS  Google Scholar 

  12. Stevens MM, George JH (2005) Exploring and engineering the cell surface interface. Science 310:1135–1138

    Article  CAS  Google Scholar 

  13. Malafaya PB, Silva GA, Reis RL (2007) Natural–origin polymers as carriers and scaffolds for biomolecules and cell delivery in tissue engineering applications. Adv Drug Deliv Rev 59:207–233

    Article  CAS  Google Scholar 

  14. Van Vlierberghe S, Dubruel P, Schacht E (2011) Biopolymer-based hydrogels as scaffolds for tissue engineering applications: a review. Biomacromolecules 12:1387–1408

    Article  Google Scholar 

  15. Martin Y, Vermette P (2005) Bioreactors for tissue mass culture: design, characterization, and recent advances. Biomaterials 26:7481–7503

    Article  CAS  Google Scholar 

  16. Chaudhuri J, Al-Rubeai M (2005) Bioreactors for tissue engineering. Springer, Dordrecht, The Netherlands

    Book  Google Scholar 

  17. Bilodeau K, Mantovani D (2006) Bioreactors for tissue engineering: focus on mechanical constraints. A comparative review. Tissue Eng 12:2367–2383

    Article  CAS  Google Scholar 

  18. Zhao J, Griffin M, Cai J, Li S, Bulter PE, Kalaskar DM (2016) Bioreactors for tissue engineering: an update. Biochem Eng J 109:268–281

    Article  CAS  Google Scholar 

  19. Mallick SP, Pal K, Rastogi A, Srivastava P (2016) Evaluation of poly (l-lactide) and chitosan composite scaffolds for cartilage tissue regeneration. Des Monomers Polym 19:271–282

    Article  CAS  Google Scholar 

  20. Jiang T, Abdel-Fattah WI, Laurencin CT (2006) In vitro evaluation of chitosan/poly (lactic acid-glycolic acid) sintered microsphere scaffolds for bone tissue engineering. Biomaterials 27:4894–4903

    Article  CAS  Google Scholar 

  21. Berninger MT, Wexel G, Rummeny EJ, Imhoff AB, Anton M, Henning TD, Vogt S (2013) Treatment of osteochondral defects in the rabbit’s knee joint by implantation of allogeneic mesenchymal stem cells in fibrin clots. JoVE (J Vis Exp) 75:e4423–e4423

    Google Scholar 

  22. Thornton G, Johnson J, Maser R, Marchuk L, Shrive N, Frank C (2005) Strength of medial structures of the knee joint are decreased by isolated injury to the medial collateral ligament and subsequent joint immobilization. J Orthop Res 23:1191–1198

    Article  CAS  Google Scholar 

  23. Rastogi A, Srivastava P, Iqbal Z, Kumaraswamy V, Singh RP (2013) Role of autologous chondrocyte transplantation in articular cartilage defects: an experimental study. Indian J Orthop 47:129

    Article  Google Scholar 

  24. Patil H, Chandel IS, Rastogi AK, Srivastava P (2013) Studies on a novel bioreactor design for chondrocyte culture. Int J Tissue Eng 2013:1–7

    Article  Google Scholar 

  25. Lao L, Tan H, Wang Y, Gao C (2008) Chitosan modified poly (l-lactide) microspheres as cell microcarriers for cartilage tissue engineering. Colloids Surf B 66:218–225

    Article  CAS  Google Scholar 

  26. Yang Q, Guo S, Wang S, Qian Y, Tai H, Chen Z (2015) Advanced glycation end products-induced chondrocyte apoptosis through mitochondrial dysfunction in cultured rabbit chondrocyte. Fundam Clin Pharmacol 29:54–61

    Article  CAS  Google Scholar 

  27. Murphy CM, Haugh MG, O’Brien FJ (2010) The effect of mean pore size on cell attachment, proliferation and migration in collagen–glycosaminoglycan scaffolds for bone tissue engineering. Biomaterials 31:461–466

    Article  CAS  Google Scholar 

  28. Bueno EM, Bilgen B, Carrier RL, Barabino GA (2004) Increased rate of chondrocyte aggregation in a wavy-walled bioreactor. Biotechnol Bioeng 88:767–777

    Article  CAS  Google Scholar 

  29. Bilgen B, Chang-Mateu IM, Barabino GA (2005) Characterization of mixing in a novel wavy-walled bioreactor for tissue engineering. Biotechnol Bioeng 92:907–919

    Article  CAS  Google Scholar 

  30. Suh J-KF, Matthew HW (2000) Application of chitosan-based polysaccharide biomaterials in cartilage tissue engineering: a review. Biomaterials 21:2589–2598

    Article  CAS  Google Scholar 

  31. Karageorgiou V, Kaplan D (2005) Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials 26:5474–5491

    Article  CAS  Google Scholar 

  32. Dash M, Chiellini F, Ottenbrite R, Chiellini E (2011) Chitosan—a versatile semi-synthetic polymer in biomedical applications. Prog Polym Sci 36:981–1014

    Article  CAS  Google Scholar 

  33. Archana D, Upadhyay L, Tewari R, Dutta J, Huang Y, Dutta P (2013) Chitosan-pectin-alginate as a novel scaffold for tissue engineering applications. Indian J. Biotechnol 12:475–482

    CAS  Google Scholar 

  34. Elsaesser A, Schwarz S, Joos H, Koerber L, Brenner R, Rotter N (2016) Characterization of a migrative subpopulation of adult human nasoseptal chondrocytes with progenitor cell features and their potential for in vivo cartilage regeneration strategies. Cell & bioscience 6:1

    Article  Google Scholar 

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Acknowledgements

This study was supported by the School of Biochemical Engineering, IIT (BHU), Department of Orthopedics, IMS, BHU, Animal House (BHU), Department of Metallurgical Engineering, IIT (BHU) and Department of Biotechnology and Medical Engineering, NIT Rourkela. We thank Dr. Vikas Singh, Dr. Abhimanyu Madhual and Mr. Dhiraj Kumar Choudhary for their assistance. The authors thanks the Indian Institute of Technology (BHU), Varanasi, India, for providing infrastructure for the research work.

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Authors and Affiliations

  1. School of Biochemical Engineering, Indian Institute of Technology (Banaras Hindu University), Varanasi, 221005, India

    Sarada Prasanna Mallick, Satyavrat Tripathi & Pradeep Srivastava

  2. Department of Orthopaedics, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005, India

    Amit Rastogi

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  1. Sarada Prasanna Mallick
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  2. Amit Rastogi
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  3. Satyavrat Tripathi
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  4. Pradeep Srivastava
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Correspondence to Pradeep Srivastava.

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Mallick, S.P., Rastogi, A., Tripathi, S. et al. Strategies on process engineering of chondrocyte culture for cartilage tissue regeneration. Bioprocess Biosyst Eng 40, 601–610 (2017). https://doi.org/10.1007/s00449-016-1724-4

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  • DOI: https://doi.org/10.1007/s00449-016-1724-4

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