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Agili-C implant promotes the regenerative capacity of articular cartilage defects in an ex vivo model

  • Susan Chubinskaya
  • Berardo Di Matteo
  • Laura Lovato
  • Francesco Iacono
  • Dror Robinson
  • Elizaveta Kon
EXPERIMENTAL STUDY

Abstract

Purpose

Osteochondral implants are currently adopted for the treatment of symptomatic full-thickness chondral and osteochondral defects. Agili-C™ is a cell-free aragonite-based scaffold which aims to reproduce the original structure and function of the articular joint while directing the growth and regeneration of both cartilage and its underlying subchondral bone. The goal of the present study was to investigate the ex vivo mechanisms of action (MOA) of the Agili-C™ implant in the repair of full-thickness cartilage defects. In particular, we tested whether Agili-C™ implant has the potential to stimulate cartilage ingrowth through chondrocytes migration into the 3D interconnected porous structure of the scaffold, along with maintaining their viability and phenotype and the deposition of hyaline cartilage matrix.

Methods

Articular cartilage samples were collected through the Gift of Hope Organ and Tissue Donor Network (Itasca, IL) within 24 h from death. For this study, cartilage from a total of 14 donors was used. To model a chondral defect, donut-shaped cartilage explants were prepared from each tissue specimen. The chondral phase of the Agili-C™ implant was placed inside the tissue in full contact and press fit manner. Cartilage explants with the Agili-C™ implant inside were cultured for 60 days. As a control, the same donut-shaped cartilage explants were cultured without Agili-C™, under the same culture conditions.

Results

Using fresh human cadaveric articular cartilage tissue in a 60-day culture, it was demonstrated that chondrocytes were able to migrate into the Agili-C™ scaffold and contribute to the deposition of the extracellular matrix (ECM) rich in collagen type II and aggrecan, and lacking collagen type I. Additionally, we were able to show the formation of a layer populated by progenitor-like cells on the articular surface of the implant.

Conclusions

The analysis of samples taken from knee and ankle joints of human donors with a wide age range and both genders supports the potential of Agili-C™ scaffold to stimulate cartilage regeneration and repair. Based on these results, the present scaffold can be used in the clinical practice as a one-step procedure to treat full-thickness chondral defects.

Keywords

Chondral defect Scaffold Biomaterial Aragonite Cartilage regeneration Ex vivo study 

Notes

Acknowledgements

The authors would like to acknowledge the Gift of Hope Organ and Tissue Donor Network and donor’s family for human tissue donation. We also would like to acknowledge the personnel from the laboratory of Dr. Chubinskaya: Dr.Arkady Margulis, MD, for tissue procurement, Dr. Lev Rappoport, MD, for histological assessment, Mrs. Arnavaz Hakimiyan for the work on the entire project. We also would like to thank Dr.Alessandra Nannini, MD for her help in manuscript editing. The project was supported by research funding from CartiHeal, LLT.

Funding

The present ex-vivo study was supported by research funding from CartiHeal, LLT.

Compliance with ethical standards

Conflict of interest

S Chubinskaya, B Di Matteo, L Lovato and F Iacono have nothing to disclose. D Robinson is an employee of CartiHeal LLT (Isreal). E Kon is a paid consultant for CartiHeal LLT (Israel).

Ethical approval

Rush University Institutional Review Board granted approval for research with human tissue specimens from deceased donors (Approval number: FWA# 00000482). The present study was conducted under the “Deceased Subjects Rule” and thus did not require the approval of other Ethical committees.

References

  1. 1.
    Glyn-Jones S, Palmer AJ, Agricola R, Price AJ, Vincent TL, Weinans H, Carr AJ (2015) Osteoarthr Lancet 386(9991):376–387CrossRefGoogle Scholar
  2. 2.
    Einhorn T, O’Keefe R, Chu C, Jacobs JJ (2013) American Academy of Orthopaedic Surgeons. Rosemont Ill Chap 10:183–198Google Scholar
  3. 3.
    Perdisa F, Filardo G, Di Matteo B, Marcacci M, Kon E (2014) Platelet rich plasma: a valid augmentation for cartilage scaffolds? A systematic review. Histol Histopathol 29(7):805–814PubMedGoogle Scholar
  4. 4.
    Kon E, Filardo G, Shani J, Altschuler N, Levy A, Zaslav K, Eisman JE, Robinson D (2015) Osteochondral regeneration with a novel aragonite-hyaluronate biphasic scaffold: up to 12-month follow-up study in a goat model. J Orthop Surg Res 10:81CrossRefGoogle Scholar
  5. 5.
    Kon E, Robinson D, Verdonk P, Drobnic M, Patrascu JM, Dulic O, Gavrilovic G, Filardo G (2016) A novel aragonite-based scaffold for osteochondral regeneration: early experience on human implants and technical developments. Injury 47(6):S27–S32CrossRefGoogle Scholar
  6. 6.
    Hattori S, Oxford C, Reddi AH (2007) Identification of superficial zone articular chondrocyte stem/progenitor cells. Biochem Biophys Res Commun 358:99–103CrossRefGoogle Scholar
  7. 7.
    Dowthwaite GP, Bishop JC, Redman SN, Khan IM, Rooney P, Evans DJ, Haughton L, Bayram Z, Boyer S, Thomson B, Wolfe MS, Archer CW (2004) The surface of articular cartilage contains a progenitor cell population. J Cell Sci 117(Pt 6):889–897CrossRefGoogle Scholar
  8. 8.
    Grogan SP, Miyaki S, Asahara H, D’Lima DD, Lotz MK (2009) Mesenchymal progenitor cell markers in human articular cartilage: normal distribution and changes in osteoarthritis. Arthritis Res Ther 11:R85CrossRefGoogle Scholar
  9. 9.
    Karlsson C, Lindahl A (2009) Articular cartilage stem cells signaling. Arthritis Res Therapy 11:121CrossRefGoogle Scholar
  10. 10.
    Bos PK, Kops N, Verhaar JA et al (2008) Cellular origin of neocartilage formed at wound edges of articular cartilage in a tissue culture experiment. Osteoarthr Cartil 16:204–211CrossRefGoogle Scholar
  11. 11.
    Chandrasekhar S, Esterman MA, Hoffman HA (1987) Microdetermination of proteoglycans and glycosaminoglycans in the presence of guanidine hydrochloride. Anal Biochem 161:103–108CrossRefGoogle Scholar
  12. 12.
    Petit B, Masuda K, D’Souza AL, Otten L, Pietryla D, Hartmann DJ et al (1996) Characterization of crosslinked collagens synthesized by mature articular chondrocytes cultured in alginate beads: comparison of two distinct matrix compartments. Exp Cell Res 225:151–161CrossRefGoogle Scholar
  13. 13.
    Masuda K, Shirota H, Thonar EJ (1994) Quantification of 35S-labeled proteoglycans complexed to alcian blue by rapid filtration in multiwell plates. Anal Biochem 217:167–175CrossRefGoogle Scholar
  14. 14.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real time quantitative PCT and the 2−∆∆CT method. Methods 25:402–408CrossRefGoogle Scholar
  15. 15.
    Calloni R, Cordero EAA, Henriques J-A-P, Bonatto D (2013) Reviewing and updating the major molecular markers for stem cells. Stem Cells Dev 22(9):1455–1476CrossRefGoogle Scholar
  16. 16.
    Cavallo C, Schiavinato A, Secchieri C, Kon E, Filardo G, Paro M, Grigolo B (2018) Short-term homing of hyaluronan-primed cells: therapeutic implications for osteoarthritis treatment. Tissue Eng Part C Methods 24(2):121–133 , CrossRefGoogle Scholar
  17. 17.
    Kon E, Filardo G, Robinson D, Eisman JA, Levy A, Zaslav K, Shani J, Altschuler N (2014) Osteochondral regeneration using a novel aragonite-hyaluronate bi-phasic scaffold in a goat model. Knee Surg Sports Traumatol Arthrosc 22(6):1452–1464CrossRefGoogle Scholar
  18. 18.
    Ko IK, Lee SJ, Atala A, Yoo JJ (2013) In situ tissue regeneration through host stem cell recruitment. Exp Mol Med 45(11):e57CrossRefGoogle Scholar
  19. 19.
    Sofu H, Camurcu Y, Ucpunar H, Ozcan S, Yurten H, Sahin V (2018) Clinical and radiographic outcomes of chitosan-glycerol phosphate/blood implant are similar with hyaluronic acid-based cell-free scaffold in the treatment of focal osteochondral lesions of the knee joint. Knee Surg Sports Traumatol Arthrosc.  https://doi.org/10.1007/s00167-018-5079-z CrossRefPubMedGoogle Scholar
  20. 20.
    Dhollander AAM, Liekens K, Almqvist KF, Verdonk R, Lambrecht S, Elewaut D, Verbruggen G, Verdonk PC (2012) A pilot study of the use of an osteochondral scaffold plug for cartilage repair in the knee and how to deal with early clinical failures. Arthroscopy 28(2):225–233CrossRefGoogle Scholar
  21. 21.
    Kon E, Delcogliano M, Filardo G, Fini M, Giavaresi G, Francioli S, Martin I, Pressato D, Arcangeli E, Quarto R, Sandri M, Marcacci M (2010) Orderly osteochondral regeneration in a sheep model using a novel nano-composite multilayered biomaterial. J Orthop Res 28(1):116–124PubMedGoogle Scholar
  22. 22.
    Verhaegen J, Clockaerts S, Van Osch GJ, Somville J, Verdonk P, Mertens P (2015) TruFit plug for repair of osteochondral defects—where is the evidence? Systematic review of literature. Cartilage 6(1):12–19CrossRefGoogle Scholar
  23. 23.
    Joshi N, Reverte-Vinaixa M, Diaz-Ferreiro EW, Dominguez Oronoz R (2012) Synthetic resorbable scaffolds for the treatment of isolated patellofemoral cartilage defects in young patients: magnetic resonance imaging and clinical evaluation. Am J Sports Med 40(6):1289–1295CrossRefGoogle Scholar
  24. 24.
    Mathis DT, Kaelin R, Rasch H, Arnold MP, Hirschmann MT (2018) Good clinical results but moderate osseointegration and defect filling of a cell-free multi-layered nano-composite scaffold for treatment of osteochondral lesions of the knee. Knee Surg Sports Traumatol Arthrosc 26(4):1273–1280PubMedGoogle Scholar
  25. 25.
    Chubinskaya S, Malfait AM, Wimmer MA (2013) Form and function of articular cartilage. In: Einhorn T, O'Keefe R, Chu C, Jacobs JJ (eds) Orthopaedic basic science: foundation of clinical practice, 4th edn. American Academy of Orthopaedic Surgeons, Rosemont, Illinois, pp 183–198Google Scholar
  26. 26.
    Angele P, Yoo JU, Smith C, Mansour J, Jepsen KJ, Nerlich M et al (2003) Cyclic hydrostatic pressure enhances the chondrogenic phenotype of human mesenchymal progenitor cells differentiated in vitro. J Orthop Res 21:451–457CrossRefGoogle Scholar
  27. 27.
    Heyland J, Wiegandt K, Goepfert C, Nagel-Heyer S, Ilinich E, Schumacher U, Pörtner R (2006) Redifferentiation of chondrocytes and cartilage formation under intermittent hydrostatic pressure. Biotechnol Lett 28(20):1641–1648CrossRefGoogle Scholar
  28. 28.
    Huang CY, Hagar KL, Frost LE, Sun Y, Cheung HS (2004) Effects of cyclic compressive loading on chondrogenesis of rabbit bone-marrow derived mesenchymal stem cells. Stem Cells 22:313–323CrossRefGoogle Scholar
  29. 29.
    Saha S, Ji L, de Pablo JJ, Palecek SP (2006) Inhibition of human embryonic stem cell differentiation by mechanical strain. J Cell Physiol 206:126–137CrossRefGoogle Scholar
  30. 30.
    Schumann D, Kujat R, Nerlich M, Angele P (2006) Mechanobiological conditioning of stem cells for cartilage tissue engineering. Biomed Mater Eng 16:S37–S52PubMedGoogle Scholar
  31. 31.
    Horbert V, Xin L, Foehr P, Brinkmann O, Bungartz M, Burgkart RH, Graeve T, Kinne RW (2018) In vitro analysis of cartilage regeneration using a collagen type I hydrogel (CaReS) in the bovine cartilage punch model. Cartilage 1:18Google Scholar
  32. 32.
    Dang Y, Cole AA, Homandberg GA (2003) Comparison of the catabolic effects of fibronectin fragments in human knee and ankle cartilages. Osteoarthr Cartil 11(7):538–547CrossRefGoogle Scholar
  33. 33.
    Eger W, Schumacher BL, Mollenhauer J, Kuettner KE, Cole AA (2002) Human knee and ankle cartilage explants: catabolic differences. J Orthop Res 20(3):526–534CrossRefGoogle Scholar

Copyright information

© European Society of Sports Traumatology, Knee Surgery, Arthroscopy (ESSKA) 2018

Authors and Affiliations

  1. 1.Rush UniversityChicagoUSA
  2. 2.Department of Biomedical SciencesHumanitas UniversityRozzano, MilanItaly
  3. 3.Humanitas Clinical and Research CenterMilanItaly
  4. 4.Rabin Medical CenterPetah TikwaIsrael
  5. 5.Sackler School of MedicineTel AvivIsrael

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