Background/Purpose:

Nerve growth factor (NGF) is a key regulator of pain and anti-NGF therapy reduces osteoarthritis (OA) associated pain. However, anti-NGF therapy is associated with rapidly progressive OA (RPOA) [1]. In hip OA there is a 5-fold increase in mesenchymal stem cells (MSCs) from MRI bone marrow (BM) lesions, areas associated with OA progression [2]. MSCs in such lesions are uniformly positive for the NGF receptor, p75, which is also linked to chemotaxis and proliferation in other stromal cell compartments [3]. We therefore sought to evaluate anti-NGF treatment in monoiodoacetate (MIA) induced OA and test whether NGF influences human BM- MSC function.

Methods:

Human tibial plateau (TP) bone was isolated from patients undergoing total knee replacement. OA was induced in male Wistar rats (n=6) by intra-articular injection of 0.3 mg MIA. Each animal was treated with subcutaneous injection of control human IgG or anti-NGF (3 mg/kg) on Days 30, 35, 39, 45 and 50. Human and animal tissues sections were prepared for histological analysis using H&E staining and immunohistochemistry (IHC) using anti-p75 and anti-NGF antibodies. BM-MSCs were isolated from iliac crest aspirates and cultured under normal conditions. Expression of p75 was induced following overnight incubation with 400mM ethanol (EtOH) and confirmed by flow cytometry. Proliferation and chemotaxis was assessed with and without induction of p75 and in the presence of 0-1 μg/ml NGF.

Results:

Regions adjacent to cartilage loss in human TP showed abundant stromal proliferation, NGF and p75 immunoreactivity. In the MIA model, by Day 28 there was substantial loss of cartilage, bone remodelling and stromal proliferation mimicking human disease and animals demonstrated unequal weight-bearing (p<0.05). Anti-NGF provided sufficient analgesia to normalise weight-bearing by Day 42. Features associated with joint damage and MIA treatment such as fibrogranular reactions and palisading osteoblasts were strongly p75 immunopositive. NGF positive staining was widespread in naïve and MIA-injected knees. To investigate increased p75 positivity in knee OA, we restored p75 expression loss in expanded cell, to 97% of BM-MSCs (n=3) using EtOH. Increased MSC proliferation was seen at Days 6 and 9 (26%, p=0.03 and 30%, p=0.01 increase respectively, n=7) for 1 μg/ml NGF in the presence of EtOH compared to control (no NGF). In cultures without EtOH induction (absent p75) NGF had no effect. NGF was not chemotactic for MSCs with or without the induction of p75.

Conclusion:

TP bone from OA patients and rat MIA-treated subchondral bone contains p75 positive staining in regions of cartilage destruction and associated NGF positivity. In vitro, NGF increased BM-MSC proliferation, suggesting NGF may be involved in the stromal proliferation seen at sites of OA damage. Thus, NGF may regulate MSC function. Complete blockade represents a novel mechanism for accelerated joint destruction in OA.

Reference:

1. Hochberg MC et al. Arthritis Rheumatol 2016;68:382–91. 2.Campbell TM et al Arthritis Rheumatol 2016 Jul;68(7):1648-59. 3. Jiang Y, Hu C, Yu S, et al. Cartilag Arthritis Res Ther 2015;:1–13.

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ACR Meeting Abstracts - https://acrabstracts.org/abstract/convergence-of-joint-repair-and-pain-pathways-via-nerve-growth-factor-and-p75-expressing-mesenchymal-stem-cells-in-established-osteoarthritis/