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Compression test on porous polymeric scaffolds

A.Parrilli1,2, F.Salamanna1, F.Veronesi1, L.Ambrosio3, A.Salerno4,5, M.Fini1 Surgical Preclinical Studies Department, Rizzoli Orthopaedic Institute, Bologna, Italy Samo S.p.A., Granarolo E. Bologna, Italy 3 Institute of Composite and Biomedical Materials and National Research Council, IMCBCNR, University of Naples Federico II, Napoli, Italy 4 Interdisciplinary Research Centre on Biomaterials, CRIB, University of Naples Federico II, Napoli, Italy 5 Italian Institute of Technology, IIT, University of Naples Federico II, Napoli, Italy
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Aims. Micro-CT is an important and valid non-destructive method, especially for the preclinical evaluations of implant materials. It is also extremely helpful when it comes to characterizing devices in the pre-implant phase and evaluating possible deformations and/or degradations in the explant phase. It is therefore useful for the analysis of both tissue engineering scaffolds and bone tissue regeneration after the preclinical application of innovative biomaterials. In this paper we aim to summarize a study on the porosity of polymeric scaffolds. Two micro-tomographies were carried out on two cylindrical polymeric scaffolds, one made of just polymer and the other one made of the same polymer with hydroxyapatite (HA) added to it. Both materials were then submitted to a compression test and analyzed through micro-CT at different levels of displacement (1mm, 3mm and maximal compression) to evaluate their changes in porosity. The two kinds of scaffolds, viewed as simple materials in this analysis, will later be used to repair a bone defect in a preclinical study in order to evaluate bone growth at different experimental times. Methods. The first step was to carry out an acquisition of the two scaffolds with the microCT system SkyScan 1172. The acquisition of both scaffolds (size is 5mm in diameter and 5mm in height) was made without using any metallic filter between the X-ray and the sample, a 167A source current and a 60kV source voltage. Both samples were rotated until 360 degrees with a rotation step of 0.30 degrees and a frame averaging of 3. The pixel size was 3.60m. The images obtained from acquisition were later reconstructed by the software NRecon (version 1.4.4) without any correction but the specific post-alignment depended on acquisition and undersampling 1. The images datasets were analyzed by CTAn (version 1.7) to evaluate total porosity, macroporosity and microporosity. In order to do that we considered a cylindrical volume of interest (VOI) totally enclosed in the scaffold, taken excluding lateral edges and some slices on the top and on the bottom of the scaffold. Then we binarized the images considering the polymeric plot as the object (white). After binarization we calculated the total porosity with the formula:

1 N

(T . Ari Obj. Ari )

T . Ari

where T.Ar is total Region of Interest Area and Obj.Ar is object Area.

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We also checked the 2D distributions of pores and the number of pores throughout the entire volume of the sample. Using these data we made a count of macro and micro porosity for each scaffold. Then we carried out the mechanical test of compression of both scaffolds inside the micro-tomograph SkyScan 1172: the first polymeric material was positioned between the two plates of the compression chamber (Figure 1) and we started by applying a load. After a displacement of 1mm, 3mm and the maximal compression, corresponding to 222N, we temporarily interrupted the compression to make new Figure 1: Compression acquisitions. chamber The mechanical test was repeated with the same method for the second sample and we recorded the graphs of load/displacement of the analyzed samples. Also in these cases the obtained datasets of reconstructed images were analyzed by CTAn to evaluate the porosity. We used the same kind of VOI totally enclosed in the scaffold. Results. The porosities of the samples before the compression test are described in table 1. Overall porosity (%) Macroporosity Microporosity POLYMER 45.3 2.5 49.7 50.3 POLYMER + HA 51.9 2.5 67.3 32.7

Table 1: Porosity of the scaffolds before the compression test Macroporosity (pore size > 50 m) has a strong impact on osteogenic outcomes and microporosity (pore size < 10 m) is believed to contribute to higher bone-inducing protein adsorption [1]. The variation of overall porosity of the polymeric scaffold submitted to the mechanical test was evaluated by plotting the percentage of porosity for each slice through the volume for every step of compression analyzed, as depicted in Figure 2. Fig ure 2: Var iati on of ove rall por osit y of the polymeric scaffold We noted a slight improvement in porosity in the first phase of compression due to an initial expansion of pores and then a progressive loss.

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We obtained nearly the same result with the polymer + hydroxyapatite, as depicted in Figure 3.

Figure 3: Variation of overall porosity of the polymeric scaffold with added hydroxyapatite The evaluation was extended to the distribution of 2D sizes of pores comparing the decrease of pore dimension in the samples after compression, as depicted in figure 4.

Figure 4: Histograms of dimension distribution of pores We noted an improvement in the microporosity after the compression, in spite of an all-in loss of porosity. The graphs of load/displacement obtained gave us information on some of the mechanical properties of the materials and we noted that adding hydroxyapatite has no significant effect on the elasticity of the polymeric scaffold. The graphs are shown in Figure 5.

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Figure 5: Load/displacement graphs Conclusion. Porous scaffolds are created with the aim of obtaining a mechanical connection, which is useful for bone growth, and which can support the stability of the implant [2]. The porosity level of a material could influence the bioactivity of the material because of the structures permeability [3]. For these reasons, it is very important to know the behaviour of the scaffold and the changes in porosity for preclinical studies. Using a Micro-CT system is extremely helpful to widen the knowledge on materials and bone response.

References [1] Karageorgiou V., Kaplan D. Porosity of 3D biomaterial scaffold and osteogenesis. Biomaterials 26 (2005); 5474-5491 [2] Puleo D.A., Nanci A. Understanding and controlling the bone-implant interface. Biomaterials. 1999 Dec;20(23-24):2311-21. Review. [3] Hing K.A. Bone repair in the twenty-first century: biology, chemistry or engineering? Philos Transact A Math Phys Eng Sci. 2004 Dec 15;362(1825):2821-50. Review.