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. 2022 Mar;40(3):573-583.
doi: 10.1002/jor.25056. Epub 2021 May 5.

Terminal sterilization influences the efficacy of an extracellular matrix-blood composite for treating posttraumatic osteoarthritis in the rat model

Naga Padmini Karamchedu  1 , Braden C Fleming  1 , Benedikt L Proffen  2 , Nicholas J Sant  2 , Gabriela Portilla  2 , Lauren R Parola  1 , Janine Molino  1 , Martha M Murray  2
Affiliations

Terminal sterilization influences the efficacy of an extracellular matrix-blood composite for treating posttraumatic osteoarthritis in the rat model

Naga Padmini Karamchedu et al. J Orthop Res. 2022 Mar.

Abstract

The objective was to determine if an intra-articular injection of an extracellular matrix (ECM) powder and blood composite (ECM-B) after anterior cruciate ligament (ACL) injury would have a mitigating effect on posttraumatic osteoarthritis and if that effect would be different with terminal sterilization of the ECM powder before use. Eighty Lewis rats underwent ACL transection and were divided into four groups: (1) intra-articular injection with phosphate-buffered saline (PBS; n = 20), (2) intra-articular injection of ECM-B using aseptically processed ECM (ASEPTIC; n = 20), (3) intra-articular injection of the ECM-busing ECM powder sterilized with 15 kGy electron beam irradiation (EBEAM; n = 20), and (4) intra-articular injection of the ECM-B using ECM powder sterilized with ethylene oxide (EO; n = 20). Twenty additional animals received capsulotomy only (SHAM). The animals were followed for 6 weeks and evaluations of gait, radiographs, and joint cartilage histology were performed. At 6 weeks, when compared to the SHAM group, the group treated with PBS had significantly worse gait and histologic changes, while the ASEPTIC group was not different from SHAM for either of these outcomes. When compared to the SHAM group, the EO group had similar gait outcomes, but greater histologic damage, and the EBEAM group had significantly worse gait and histological outcomes. The ECM-B composite produced using aseptically processed ECM powder mitigated the gait and histologic changes associated with osteoarthritis after ACL transection in the rat; however, care must be taken when selecting a terminal sterilization method as this may affect the effectiveness of treatment.

Keywords: ACL; ECM gel injection; articular cartilage; posttraumatic osteoarthritis; rat.

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Figures

Figure 1.
Figure 1.
Schematic diagram of the experimental design. ACLT=ACL Transection; Sx=Surgery; ECM-B=extracellular matrix blood composite; EO=ethylene oxide; PBS=phosphate buffered saline.
Figure 2.
Figure 2.
Mean difference in maximum force relative to baseline (%BW) at (A) 2 weeks and at (B) 6 weeks between the surgical and contralateral control knees. A negative value indicates more offloading of the surgical limb. Error bars represent the 95% confidence intervals. BW=body weight; PBS=phosphate buffered saline; EO=ethylene oxide; EBEAM=electron beam; ASEP=ASEPTIC.
Figure 3.
Figure 3.
Mean Kellgren-Lawrence scores for the knees in each treatment group. Error bars represent the 95% confidence intervals. Only the EBEAM group had significantly higher scores than the SHAM group (*P=.01). All groups had mild changes with mean scores between 0 (normal) and 1 (possible osteophytes). PBS=phosphate buffered saline; EO=ethylene oxide; EBEAM=electron beam.
Figure 4.
Figure 4.
Representative images showing the cartilage damage (black arrows) in the medial compartment of the surgical limbs for each treatment group. PBS=phosphate buffered saline; EO=ethylene oxide; EBEAM=electron beam.
Figure 5.
Figure 5.
Histological analysis of synovitis in the medial compartment of the treated knees. Synovitis was scored adjacent to the medial meniscus according to a synovial membrane inflammation score. The mean values and the 95% confidence intervals are provided. Knees that underwent a capsulotomy with no ACL transection (SHAM) had significantly less synovitis when compared to all other groups (*** P<.001 pbs="phosphate" buffered saline eo="ethylene" oxide ebeam="electron" beam.>
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