Polyetheretherketone Multifilament and Monofilament

Woven Tissue Engineering Scaffolds

1
Edwards, S.L.,
2
Werkmeister, J.A.,
2
Ramshaw, J.A.M.,
2
McLean, K.,
3
Jarman-Smith, M.L.
1
CSIRO Materials Science and Engineering, Geelong, 3216, Australia
2
CSIRO Molecular and Health Technologies, Clayton, 3164, Australia
3
Invibio Ltd., Thornton-Cleveleys, UK
contact: Sharon Edwards
phone: (61) 3 5246 4736
email: sharon.edwards@csiro.au
web: http://www.csiro.au/people/Sharon.Edwards.html
Materials and Methods
Implantable grade PEEK Optima (Invibio Ltd.; UK) monofilament (10.2 tex) and
multifilament (7.5 tex/15 filaments) yarns, and PET multifilament yarn (8.3 tex/36
filaments), were woven into plain weave scaffolds. Following standard methods
fabrics were assessed for their physical and mechanical properties.
All scaffolds were washed and sterilised (autoclaved) prior to cell seeding. Mouse
L929 fibroblast cells (passage 13) were suspended in supplemented Dulbeccos
modified Eagles medium at a concentration of 6.5 x 10
4
cells/mL. Cell suspension
(1.37 mL) was statically seeded onto scaffolds, secured using polymer ring holders.
Samples were incubated for up to 16 days in vitro, changing the medium three times
per week. Prior to conducting the assays scaffolds were rinsed in phosphate
buffered saline to remove non-adherent cells. At regular intervals an MTS assay
(CellTiter 96

, Promega) was performed to give an indication of cell attachment and

proliferation. Cell viability was assessed using a viability assay (Live/Dead

,
Molecular Probes), using fluorescent microscopy to visualise live, calcein AM
stained (green) cells and dead, ethidium homodimer-1 stained (red) cells. Scanning
electron microscopy (SEM) was used to observe cell morphology, using standard
sample preparation procedures.
Introduction
Polyetheretherketone (PEEK) is a semi-crystalline thermoplastic polymer. It
combines good strength and stiffness with excellent thermal stability and good
chemical resistance. Studies have found good biocompatibility with fibroblasts and
osteoblasts in vitro [1, 2], with no cytotoxic effects in vivo [3]. Given these properties,
PEEK has been used in a number of biomedical applications, including spinal disc
fusion, bone trauma repair, and craniomaxillofacial repair [4]. PEEK medical devices
are typically produced by manufacturing methods like injection moulding [5], laser
sintering [6], and machining. To meet the demands of more flexible implant devices,
other methods of manufacture are required.
The aim of this work was to fabricate flexible woven PEEK scaffolds. Scaffolds were
characterised in terms of their physical and mechanical properties and biologically
assessed, culturing with L929 mouse fibroblasts for up to 16 days in vitro. Results
were compared to those obtained for fabricated polyethylene terephthalate (PET)
woven scaffolds.
a
References
1. Sagomonyants KB. Biomaterials. 2008;29: 1563-1572.
2. Hunter A. Biomaterials. 1995;16:287-295.
3. Rivard CH. J Biomed Mater Res. 2002;62:488-498.
4. Jarman-Smith ML. Medical Device Tech. 2008;19:12-15.
5. Nieminen T. J Biomed Mater Res. 2008;84A:377-383.
6. Tan KH, Bio-Med Mater and Eng. 2005;15:113-124.
Conclusions
Multifilament PEEK woven scaffolds were lighter, thinner and stronger than
monofilament PEEK scaffolds. Woven PEEK scaffolds supported fibroblast cell
attachment and proliferation, and ECM production. Cells were found to orientate with
the filament direction, and in places, span the inter-filament and inter-yarn pores of the
multifilament scaffold. The results of this study indicate that fibrous PEEK structures
have potential as scaffolds for tissue engineering.
Results
The multifilament PEEK scaffold was found to be of lower mass/area, thinner, with
smaller inter-yarn pores, compared to the PEEK monofilament scaffold. This was due
to the multifilament yarn possessing a lower linear density (mass per unit length), and
smaller filament diameter. The smaller inter-yarn pores were a consequence of yarn
flattening within the woven structure. The multifilament scaffold was found to be
stronger and slightly less extensible (in both directions) than the monofilament
scaffold, with higher burst strength. See Table 1.
Monofilament PEEK
scaffold
Multifilament PEEK
scaffold
Mass/area (g/m
2
) 74.82 (1.03) 55.03 (0.44)
Thickness (mm) 0.69 (0.01) 0.18 (0.01)
Max. pore size (m) 336.75 (4.84) 111.01 (8.60)
Burst Strength (N) 401 492
Tensile load at break (N)
534L (14.48)
366W (38.74)
657L (51.36)
548W (14.43)
Tensile strain at break (%) 40L (0.89) 21W (1.63) 18L (1.81) 18W (1.05)
Table 1: Properties of fabricated PEEK scaffolds.
Results expressed as mean (standard error of the mean).
L denotes warp direction and W denotes weft direction.
SEM micrographs revealed cells to be flattened with processes extending onto the
PEEK substrate (Figure 2a), with extracellular matrix (ECM) deposition. Cells were
observed to span across neighbouring filaments (Figure 2b) and partially infiltrate the
inter-yarn pores (Figure 2c) of the multifilament scaffolds. Conversely, the larger inter-
yarn distances of the monofilament scaffold did not appear to permit cell spanning.
See Figure 2d.
All scaffolds were found to support cell attachment, with proliferation up to day 8 of
culture. The MTS assay determined the PET multifilament scaffold to sustain slightly
higher cell numbers in the early stages of culture. This was likely due to smaller inter-
yarn pores (average of 73 m), which may have resulted in a higher cell seeding
efficiency, and a higher scaffold surface area for cell growth. With increased
incubation time similar cell numbers were found on all scaffolds, with slightly lower
numbers on the PEEK monofilament scaffold by day 10 of culture.
Figure 1: Fluorescent micrographs of viable (green) and dead (red) fibroblasts on (a)
PEEK monofilament, (b) PEEK multifilament, and (c and d) PET multifilament
scaffolds on days (a-c) 6 and (d) 10 of culture.
a
d c
b
The viability assay revealed few dead cells on the scaffolds throughout the culture
period. Little infiltration into the large pores of the monofilament PEEK scaffold was
observed by day 10. Conversely, the smaller pores of the multifilament scaffold
were partially filled (Figures 1a-b). Cells were found to be aligned with the filament
direction on the multifilament scaffolds (Figures 1c-d), but not the monofilament
scaffolds.
Figure 2: SEM micrograph of fibroblasts on (a and d) PEEK monofilament scaffold
and (b and c) PEEK multifilament scaffold, on day 8 of culture.
b a
d c