Skip to main content
SpringerLink
Log in

In vitro performance of injectable chitosan-tripolyphosphate scaffolds combined with platelet-rich plasma

  • Original Article
  • Published:
Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

This study aimed to evaluate the in vitro biological effectiveness of chitosan microparticles crosslinked with sodium tripolyphosphate (TPP) in combination with activated pure platelet-rich plasma (aP-PRP) as an injectable composite scaffold for growth factors release, cell proliferation and osteogenic differentiation. Two main novelties were addressed in the field of scaffolds in regenerative medicine: the first is the approach including simultaneously the three vertices of the proliferation triangle formed by the capabilities genic progenitor cells, conductive scaffolds and inductive growth factors, which are provided by platelet rich plasma (PRP); secondly, the approach of an injectable composite scaffolds formed by the fibrin network from aP-PRP and the chitosan microparticles crosslinked with TPP. The microparticles were prepared by vortexing the chitosan and TPP solutions. The ionic crosslinking of chitosan with TPP was made at mass ratios of 2:1, 5:1, and 10:1 at pH 4.0. P-PRP was obtained via the controlled centrifugation of whole blood. The composite scaffolds were prepared by adding the microparticles to immediately activated P-PRP. The results showed that the microparticles had adequate physicochemical and mechanical properties for injection. Furthermore, the microparticles controlled the release of growth factors from P-PRP. The proliferation of human adipose-derived mesenchymal stem cells was lower than in aP-PRP alone but significant at a 2:1 chitosan-TPP mass ratio. Osteogenic differentiation was stimulated at all studied mass ratios, as indicated by the alkaline phosphatase activity. These results offer perspectives for optimizing the composite scaffold, and to prove its potential as an injectable scaffold in regenerative medicine.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Kretlow JD, Klouda L, Mikos AG. Injectable matrices and scaffolds for drug delivery in tissue engineering. Adv Drug Deliv Rev 2007;59:263–273.

    Article  CAS  PubMed  Google Scholar 

  2. Hou Q, De Bank PA, Shakesheff KM. Injectable scaffolds for tissue regeneration. J Mater Chem 2004;14:1915–1923.

    Article  CAS  Google Scholar 

  3. Gutowska A, Jeong B, Jasionowski M. Injectable gels for tissue engineering. Anat Rec 2001;263:342–349.

    Article  CAS  PubMed  Google Scholar 

  4. Costa-Pinto AR, Reis RL, Neves NM. Scaffolds based bone tissue engineering: the role of chitosan. Tissue Eng Part B Rev 2011;17:331–347.

    Article  CAS  PubMed  Google Scholar 

  5. Berger J, Reist M, Mayer JM, Felt O, Peppas NA, Gurny R. Structure and interactions in covalently and ionically crosslinked chitosan hydrogels for biomedical applications. Eur J Pharm Biopharm 2004;57:19–34.

    Article  CAS  PubMed  Google Scholar 

  6. Brack HP, Tirmizi SA, Risen Jr WM. A spectroscopic and viscometric study of the metal ion-induced gelation of the biopolymer chitosan. Polymer 1997;38:2351–2362.

    Article  CAS  Google Scholar 

  7. Hennink WE, van Nostrum CF. Novel crosslinking methods to design hydrogels. Adv Drug Deliv Rev 2002;54:13–36.

    Article  CAS  PubMed  Google Scholar 

  8. Bhumkar DR, Pokharkar VB. Studies on effect of pH on cross-linking of chitosan with sodium tripolyphosphate: a technical note. AAPS PharmSciTech 2006;7:E50.

    Article  PubMed  Google Scholar 

  9. Ibezim EC, Andrade CT, Marcia C, Barretto B, Odimegwu DC, de Lima FF. Ionically Cross-linked Chitosan/Tripolyphosphate Microparticles for the Controlled Delivery of Pyrimethamine. Ibnosina J Med BS 2011;3:77–88.

    Google Scholar 

  10. Harris R, Lecumberri E, Heras A. Chitosan-genipin microspheres for the controlled release of drugs: clarithromycin, tramadol and heparin. Mar Drugs 2010;8:1750–1762.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Karnchanajindanun J, Srisa-ard M, Srihanam P, Baimark Y. Preparation and characterization of genipin-cross-linked chitosan microparticles by water-in-oil emulsion solvent diffusion method. Natural Science 2010;2:1061–1065.

    Article  CAS  Google Scholar 

  12. Barnett Jr MD, Pomeroy GC. Use of platelet-rich plasma and bone marrow-derived mesenchymal stem cells in foot and ankle surgery. Tech Foot Ankle Surg 2007;6:89–94.

    Article  Google Scholar 

  13. Crane D, Everts PAM. Platelet Rich Plasma (PRP) Matrix Grafts. Pract Pain Manag 2008;8:12–26.

    Google Scholar 

  14. Foster TE, Puskas BL, Mandelbaum BR, Gerhardt MB, Rodeo SA. Platelet-rich plasma: from basic science to clinical applications. Am J Sports Med 2009;37:2259–2272.

    Article  PubMed  Google Scholar 

  15. Marx RE. Platelet-rich plasma: evidence to support its use. J Oral Maxillofac Surg 2004;62:489–496.

    Article  PubMed  Google Scholar 

  16. Tabata Y. Tissue regeneration based on growth factor release. Tissue Eng 2003;9 Suppl 1:S5–S15.

    Article  CAS  PubMed  Google Scholar 

  17. Shimojo AA, Perez AG, Galdames SE, Brissac IC, Santana MH. Performance of PRP associated with porous chitosan as a composite scaffold for regenerative medicine. ScientificWorldJournal 2015;2015:396131.

  18. Nasti A, Zaki NM, de Leonardis P, Ungphaiboon S, Sansongsak P, Rimoli MG, et al. Chitosan/TPP and chitosan/TPP-hyaluronic acid nanoparticles: systematic optimisation of the preparative process and preliminary biological evaluation. Pharm Res 2009;26:1918–1930.

    Article  CAS  PubMed  Google Scholar 

  19. Flory PJ, Rehner Jr J. Statistical Mechanics of Cross-Linked Polymer Networks I. Rubberlike Elasticity. J Chem Phys 1943;11:512–520.

    Article  CAS  Google Scholar 

  20. Collins MN, Birkinshaw C. Investigation of the swelling behavior of crosslinked hyaluronic acid films and hydrogels produced using homogeneous reactions. J Appl Polym Sci 2008;109:923–931.

    Article  CAS  Google Scholar 

  21. Jin J, Song M. Chitosan and chitosan-PEO blend membranes crosslinked by genipin for drug release. J Appl Polym Sci 2006;102:436–444.

    Article  CAS  Google Scholar 

  22. Rädler JO, Koltover I, Jamieson A, Salditt T, Safinya CR. Structure and interfacial aspects of self-assembled cationic lipid-DNA gene carrier complexes. Langmuir 1998;14:4272–4283.

    Article  Google Scholar 

  23. Tan H, Chu CR, Payne KA, Marra KG. Injectable in situ forming biodegradable chitosan-hyaluronic acid based hydrogels for cartilage tissue engineering. Biomaterials 2009;30:2499–2506.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55–63.

    Article  CAS  PubMed  Google Scholar 

  25. Gümüsderelioglu M, Aday S. Heparin-functionalized chitosan scaffolds for bone tissue engineering. Carbohydr Res 2011;346:606–613.

    Article  PubMed  Google Scholar 

  26. Perez AG, Lichy R, Lana JF, Rodrigues AA, Luzo AC, Belangero WD, et al. Prediction and modulation of platelet recovery by discontinuous centrifugation of whole blood for the preparation of pure platelet-rich plasma. Biores Open Access 2013;2:307–314.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Mi FL, Shyu SS, Lee ST, Wong TB. Kinetic study of chitosan-tripolyphosphate complex reaction and acid-resistive properties of the chitosan-tripolyphosphate gel beads prepared by in-liquid curing method. J Polym Sci Pol Phys 1999;37:1551–1564.

    Article  CAS  Google Scholar 

  28. Kablik J, Monheit GD, Yu L, Chang G, Gershkovich J. Comparative physical properties of hyaluronic acid dermal fillers. Dermatol Surg 2009;35 Suppl 1:302–312.

    Article  CAS  PubMed  Google Scholar 

  29. Ikeda S, Nishinari K. “Weak gel”-type rheological properties of aqueous dispersions of nonaggregated kappa-carrageenan helices. J Agric Food Chem 2001;49:4436–4441.

    Article  CAS  PubMed  Google Scholar 

  30. Clark AH, Ross-Murphy SB. Structural and mechanical properties of biopolymer gels. Adv Polym Sci 1987;83:57–192.

    Article  CAS  Google Scholar 

  31. de Melo F, Marijnissen-Hofsté J. Investigation of physical properties of a polycaprolactone dermal filler when mixed with lidocaine and lidocaine/epinephrine. Dermatol Ther (Heidelb) 2012;2:13.

    Article  Google Scholar 

  32. Ohrlund A, Edsman K, Sturesson C, Nord L, Helander Kenne A, Nasstrom J. Lifting capacity of hyaluronic acid (HA) dermal fillers. Presented at: 8th Anti-Aging Medicine World Congress & MeiSpa (AMWC); 2010 Apr 8-10; Monte-Carlo, Monaco.

    Google Scholar 

  33. Bentkover SH. The biology of facial fillers. Facial Plast Surg 2009;25:73–85.

    Article  CAS  PubMed  Google Scholar 

  34. Peppas NA, Sahlin JJ. Hydrogels as mucoadhesive and bioadhesive materials: a review. Biomaterials 1996;17:1553–1561.

    Article  CAS  PubMed  Google Scholar 

  35. Hokugo A, Ozeki M, Kawakami O, Sugimoto K, Mushimoto K, Morita S, et al. Augmented bone regeneration activity of platelet-rich plasma by biodegradable gelatin hydrogel. Tissue Eng 2005;11:1224–1233.

    Article  CAS  PubMed  Google Scholar 

  36. De Cock LJ, De Wever O, Hammad H, Lambrecht BN, Vanderleyden E, Dubruel P, et al. Engineered 3D microporous gelatin scaffolds to study cell migration. Chem Commun (Camb) 2012;48:3512–3514.

    Article  Google Scholar 

  37. Wang C, Li J, Yao F. Application of Chitosan-Based Biomaterials in Tissue Engineering. In: Yuji Y, editor. Chitosan-Based Hydrogels: Functions and Applications. New York: CRC Press; 2011. p. 424.

    Google Scholar 

  38. Stein GS, Lian JB. Molecular mechanisms mediating developmental and hormone-regulated expression of genes in osteoblasts: an integrated relationship of cell growth and differentiation. In: Noda M, editor. Cellular and molecular biology of bone. Tokyo: Academic Press; 1993. p. 47–95.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Engineering of Materials and Bioprocesses, School of Chemical Engineering, University of Campinas (UNICAMP), Campinas, Brazil

    Andréa Arruda Martins Shimojo, Sofia Elisa Moraga Galdames, Amanda Gomes Marcelino Perez & Maria Helena Andrade Santana

  2. Department of Physical Chemistry, Institute of Chemistry, University of Campinas (UNICAMP), Campinas, Brazil

    Thiago Heiji Ito

  3. Haematology and Hemotherapy Center, Umbilical Cord Blood Bank, University of Campinas (UNICAMP), Campinas, Brazil

    Ângela Cristina Malheiros Luzo

Authors
  1. Andréa Arruda Martins Shimojo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sofia Elisa Moraga Galdames
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Amanda Gomes Marcelino Perez
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Thiago Heiji Ito
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ângela Cristina Malheiros Luzo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Maria Helena Andrade Santana
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Maria Helena Andrade Santana.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shimojo, A.A.M., Galdames, S.E.M., Perez, A.G.M. et al. In vitro performance of injectable chitosan-tripolyphosphate scaffolds combined with platelet-rich plasma. Tissue Eng Regen Med 13, 21–30 (2016). https://doi.org/10.1007/s13770-015-9111-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13770-015-9111-9

Keywords

Search

Navigation