Acute mobilization and migration of bone marrow-derived stem cells following anterior cruciate ligament rupture T. Maerz, M. Fleischer, M.D. Newton,

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Presentation transcript:

Acute mobilization and migration of bone marrow-derived stem cells following anterior cruciate ligament rupture T. Maerz, M. Fleischer, M.D. Newton, A. Davidson, M. Salisbury, P. Altman, M.D. Kurdziel, K. Anderson, A. Bedi, K.C. Baker Osteoarthritis and Cartilage Volume 25, Issue 8, Pages 1335-1344 (August 2017) DOI: 10.1016/j.joca.2017.03.004 Copyright © 2017 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Representative in vivo NIR images of contralateral (A and C) and injured/uninjured (B and D) limbs of both Control (A and B) and Rupture (C and D) rats at 72 h. Ruptured limbs (D) exhibited significantly higher normalized NIR signal within the ROI (dotted circle) encompassing the knee joint at all imaging time points (E). Normalized NIR signal was calculated by dividing NIR fluorescence signal intensity of the right limb (uninjured/injured) by the left limb (contralateral). ** indicates P < 0.001. Error bars represent 95% confidence interval of the mean. Results reflect n = 6 rats per group. Osteoarthritis and Cartilage 2017 25, 1335-1344DOI: (10.1016/j.joca.2017.03.004) Copyright © 2017 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Ex vivo NIR imaging was performed on major knee joint tissues (A, left-to-right, top-to-bottom: quadriceps muscle, femur. tibia, synovium, ACL, medial and lateral meniscus) as well as the major filtering organs (B, left-to-right, top-to-bottom: kidneys, liver, lungs, spleen). Normalizing the injured/uninjured limb to its contralateral, significantly higher signal was measured in the quadriceps and synovium of Rupture rats (C). Normalizing each filtering organ to naïve controls which did not receive fluorescent cell injections, the majority of fluorescent signal was detected in the lungs and long bones (D). Error bars represent 95% confidence interval of the mean. Results reflect n = 6 rats per group. * indicates P < 0.05. ** indicates P < 0.001. Osteoarthritis and Cartilage 2017 25, 1335-1344DOI: (10.1016/j.joca.2017.03.004) Copyright © 2017 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 H&E (A, C, E) and corresponding unstained fluorescent (B, D, F) histologic sections from injured limbs, demonstrating localization of fluorescently-labeled MSCs, indicated with triangular arrows. Bolt arrows indicate autofluorescent RBCs. In quadriceps sections (A, B), labeled MSCs were localized at the MTJ (M = muscle, T = tendon). In synovial sections (C, D), labeled MSCs were localized in synovial tissue in the vicinity of arterioles (A = arterioles). In femoral sections (E, F), labeled MSCs were localized within bone marrow, but not in mineralized bone or articular cartilage (AC = articular cartilage, BM = bone marrow). Osteoarthritis and Cartilage 2017 25, 1335-1344DOI: (10.1016/j.joca.2017.03.004) Copyright © 2017 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Synovial histologic sections stained with H&E (A) and IHC stains for SDF-1 (B) and interleukin 17A (IL-17A, C). H&E-stained sections from control rats appeared normal and exhibited little to no differences between uninjured (A1) and contralateral (A3) limbs, while sections from injured limbs (A2) demonstrated marked superficial hypercellularity and hyperplasticity compared to its contralateral (A4) as well as control limbs (A1,3). SDF-1 IHC sections demonstrated similar degrees of positive SDF-1 staining in control uninjured (B1), control contralateral (B2), and rupture contralateral limbs (B4). Injured limbs (B2) demonstrated the highest levels of positive SDF-1 staining, but there was considerable variability in stain intensity between rats. IL-17A IHC sections from control rats demonstrated relatively low levels of positive IL-17A staining, with no difference in positive staining between the uninjured (C1) and contralateral (C3) limbs, while sections from injured (C2) limbs demonstrate a substantially greater degree of positive IL-17A staining than its contralateral (C4) or either control limb (C1,3). Positive IL-17 staining was identified both diffusely throughout the extracellular matrix as well as intracellularly. Osteoarthritis and Cartilage 2017 25, 1335-1344DOI: (10.1016/j.joca.2017.03.004) Copyright © 2017 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Fluorescent signal from the serum cytokine panel did not indicate any significant differences in serum cytokine or chemokine concentration between Rupture and Control groups. Error bars represent 95% confidence interval of the mean. Results reflect n = 6 rats per group. Osteoarthritis and Cartilage 2017 25, 1335-1344DOI: (10.1016/j.joca.2017.03.004) Copyright © 2017 Osteoarthritis Research Society International Terms and Conditions