*Article* **Impact of Three Different Processing Techniques on the Strength and Structure of Juvenile Ovine Pulmonary Homografts**

**Johannes J van den Heever 1,\* , Christiaan J Jordaan <sup>1</sup> , Angélique Lewies <sup>1</sup> , Jacqueline Goedhals <sup>2</sup> , Dreyer Bester <sup>1</sup> , Lezelle Botes <sup>3</sup> , Pascal M Dohmen 1,4 and Francis E Smit <sup>1</sup>**


**Abstract:** Homografts are routinely stored by cryopreservation; however, donor cells and remnants contribute to immunogenicity. Although decellularization strategies can address immunogenicity, additional fixation might be required to maintain strength. This study investigated the effect of cryopreservation, decellularization, and decellularization with additional glutaraldhyde fixation on the strength and structure of ovine pulmonary homografts harvested 48 h post-mortem. Cells and cellular remnants were present for the cryopreserved group, while the decellularized groups were acellular. The decellularized group had large interfibrillar spaces in the extracellular matrix with uniform collagen distribution, while the additional fixation led to the collagen network becoming dense and compacted. The collagen of the cryopreserved group was collapsed and appeared disrupted and fractured. There were no significant differences in strength and elasticity between the groups. Compared to cryopreservation, decellularization without fixation can be considered an alternative processing technique to maintain a well-organized collagen matrix and tissue strength of homografts.

**Keywords:** homografts; ischaemic harvesting; decellularization; cryopreservation; glutaraldehyde-fixation
