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In vivo evaluation of the local inflammatory response after intramuscular implantation of Ti6Al4V plates with anti-adhesive plasma-fluorocarbon-polymer films in rats

Michael Schlosser1, Birgit Finke2, Holger Testrich2, 3, Silke Lucke1, Uwe Walschus1, Karsten Schröder2, Jürgen Meichsner3, Maciej Patrzyk4 and Andreas Hoene4
  • 1 University Medical Center Greifswald, Department of Medical Biochemistry and Molecular Biology, Germany
  • 2 Leibniz Institute for Plasma Science and Technology (INP), Germany
  • 3 Ernst Moritz Arndt University of Greifswald, Institute of Physics, Germany
  • 4 University Medical Center Greifswald, Department of Surgery, Germany

Introduction: Titanium (Ti) alloys like Ti6Al4V are often used for temporary osteosynthesis devices and other orthopaedic implants. Unlike permanent implants, temporary devices should ideally be characterized by limited cell-material contact strength to facilitate uncomplicated removal after bone healing has been accomplished. Anti-adhesive plasma-fluorocarbon-polymer (PFP) films created by low-temperature plasma processes can be used to restrict cell-material interactions[1]. The aim of the present study was to evaluate the local inflammatory tissue response following implantation of Ti6Al4V samples with different PFP films in rats.

Materials and Methods: Ti6Al4V plates (5x5x1mm) were equipped with PFP film by (1) micro-wave discharge plasma (MW) with a C3F8/ H2 or C6F6/ H2 mix or (2) radiofrequency discharge plasma (RF) with a C3F8/ H2 mix, resulting in 3 sample series designated as MW-C3F8, MW-C6F6 and RF-C3F8, respectively. Uncoated plasma-treated plates were used as controls. One sample from each PFP series and one control sample were simultaneously implanted into the neck musculature of 24 male Lewis rats. After 7, 14 and 56 days, the samples with peri-implant tissue were retrieved from 8 animals/ day. Tissue cryosections were stained with antibodies for CD68+ macrophages/ monocytes (ED1), CD163+ macrophages (ED2), T cells (R73), IL-2R+ cells (OX39), MHC-II+ antigen-presenting cells (OX6) and activated NK cells (ANK61), and  the APAAP system. Toluidine blue was used for histochemical staining of mast cells. Morphometric results were obtained using ImageJ software and expressed as percentage of stained area or cells per mm2.

Results and Discussion: The CD68+ macrophages/ monocytes response was stronger for MW-C3F8 than for controls on day 14, while MW-C6F6 caused a significantly stronger response on day 56. In contrast, RF-C3F8 did not differ from controls on any day. CD163+ macrophages had a significantly stronger response for both MW films compared to controls on day 14 with no differences on day 56, while antigen-presenting cells were similarly significantly increased for both MW films on day 14 but only for MW-C6F6 on day 56. The T cell response was increased on day 56 for all PFP series in comparison to controls, with similar results for IL-2R+ cells. Furthermore, NK and mast cells were increased for MW-C6F6 compared to controls on day 14.

Conclusion: In summary, RF-C3F8 films were comparable to controls regarding local tissue reactions. Taken together with previous in vitro results, these data demonstrate their potential for anti-adhesive coating of orthopaedic devices. The cellular response was influenced by the choice of plasma technology, precursor chemistry and process parameters, which could thus be adapted for specific applications.

The study was supported by the Federal Ministry of Education & Research in the Campus PlasmaMed (project PlasmaImp).

References:
[1] Finke B. et al., Mater Sci Forum 783-786:1238, 2014

Keywords: in vivo, Biocompatibility, Cell response, bioinerface

Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.

Presentation Type: Poster

Topic: Biomaterials evaluation in animal models

Citation: Schlosser M, Finke B, Testrich H, Lucke S, Walschus U, Schröder K, Meichsner J, Patrzyk M and Hoene A (2016). In vivo evaluation of the local inflammatory response after intramuscular implantation of Ti6Al4V plates with anti-adhesive plasma-fluorocarbon-polymer films in rats. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02839

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Received: 27 Mar 2016; Published Online: 30 Mar 2016.