Total joint replacement (TJR) is a highly successful surgical procedure;however the long-term survivorship is limited by wear of the bearing surfaces. Wear particles stimulate chronic inflammation that delays osseointegration, and leads to peri-prosthetic osteolysis and implant loosening. Particle-induced inflammation results in the release of pro-inflammatory factors;these events are mediated primarily by the transcription factor Nuclear Factor kappa B (NF?B), a critical signaling molecule in the activation of pro- inflammatory genes. The purpose of this grant is to modulate NF?B expression in order to mitigate particle- induced inflammation and osteolysis using in vitro and in vivo models. The research proposal will yield substantive strategies for treatment of periprosthetic osteolysis in humans.
Specific Aim #1 : To demonstrate that an NF?B decoy oligodeoxynucleotide (ODN) will abrogate the expression of pro-inflammatory genes and proteins when murine or human macrophages are exposed to clinically relevant polyethylene particles with/without adherent endotoxin in vitro.
Specific Aim #2 : To demonstrate that an NF?B decoy ODN will have no major adverse effects on murine or human osteoprogenitors when the cells are exposed to clinically relevant polyethylene particles with/without adherent endotoxin in vitro.
Specific Aim #3 : To demonstrate that local delivery of the NF?B decoy ODN inhibits peri-implant inflammation and osteolysis associated with continuous infusion of polyethylene particles using our validated murine femoral implant model.
Specific Aim #4 : To demonstrate that local delivery of the NF?B decoy ODN inhibits systemic migration of exogenous, reporter macrophages to the area of particle infusion using our murine model. Murine and human macrophages and osteoprogenitor cells (OPCs) will be cultured with clinically relevant polyethylene particles endotoxin. NF?B decoy ODN, scrambled decoy ODN or nothing will be added to the cultures. Cellular viability and proliferation will be assessed;TNF, IL-1, IL-6, MCP-1 and RANKL will be assayed at the protein and gene levels from the supernatants (ELISA and Western blot) and cells (RT- PCR) respectively. OPC cultures will be assayed for alkaline phosphatase and osteocalcin expression, and bone matrix formation will be quantitated using Von Kossa staining. Continuous infusion of polyethylene particles using our murine femoral implant model will result in systemic reporter macrophage trafficking to the site of particle infusion (using bioluminescence and immunofluorescence microscopy), increased local bone turnover (using microPET scans) and osteolysis (using quantitative microCT). These effects will be mitigated by the local delivery of NF?B decoy ODN. Polyethylene debris will continue to be generated from TJRs, even with newer polyethylenes that produce smaller, potentially more biologically active particles. The proposed research is important, innovative and clinically relevant;it will suggest potential local pharmacological strategies to mitigate the chronic inflammatory reaction to wear particles and periprosthetic osteolysis.

Public Health Relevance

Total joint replacement (TJR) is a highly successful surgical procedure for end-stage arthritis, however the longevity of TJRs is limited by wear of the bearing surfaces. Wear particles stimulate a chronic inflammatory reaction that leads to local bone destruction around the implant. This grant application tests a novel translational strategy to mitigate particle-associated bone destruction by interfering with the primary signaling molecule (the transcription factor NFkB) within cells that controls the production of inflammatory mediators. This strategy has a high likelihood of extending the lifetime of TJRs in humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
1R01AR063717-01
Application #
8419372
Study Section
Musculoskeletal Tissue Engineering Study Section (MTE)
Program Officer
Panagis, James S
Project Start
2012-09-15
Project End
2017-07-31
Budget Start
2012-09-15
Budget End
2013-07-31
Support Year
1
Fiscal Year
2012
Total Cost
$353,361
Indirect Cost
$128,361
Name
Stanford University
Department
Surgery
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Nabeshima, Akira; Pajarinen, Jukka; Lin, Tzu-Hua et al. (2017) Mutant CCL2 protein coating mitigates wear particle-induced bone loss in a murine continuous polyethylene infusion model. Biomaterials 117:1-9
Córdova, Luis A; Loi, Florence; Lin, Tzu-Hua et al. (2017) CCL2, CCL5, and IGF-1 participate in the immunomodulation of osteogenesis during M1/M2 transition in vitro. J Biomed Mater Res A 105:3069-3076
Gibon, Emmanuel; Córdova, Luis A; Lu, Laura et al. (2017) The biological response to orthopedic implants for joint replacement. II: Polyethylene, ceramics, PMMA, and the foreign body reaction. J Biomed Mater Res B Appl Biomater 105:1685-1691
Lin, Tzuhua; Pajarinen, Jukka; Nabeshima, Akira et al. (2017) Preconditioning of murine mesenchymal stem cells synergistically enhanced immunomodulation and osteogenesis. Stem Cell Res Ther 8:277
Lin, T-H; Pajarinen, J; Lu, L et al. (2017) NF-?B as a Therapeutic Target in Inflammatory-Associated Bone Diseases. Adv Protein Chem Struct Biol 107:117-154
Pajarinen, Jukka; Lin, Tzu-Hua; Nabeshima, Akira et al. (2017) Mesenchymal stem cells in the aseptic loosening of total joint replacements. J Biomed Mater Res A 105:1195-1207
Lin, Tzu-Hua; Gibon, Emmanuel; Loi, Florence et al. (2017) Decreased osteogenesis in mesenchymal stem cells derived from the aged mouse is associated with enhanced NF-?B activity. J Orthop Res 35:281-288
Lin, Tzuhua; Pajarinen, Jukka; Nabeshima, Akira et al. (2017) Establishment of NF-?B sensing and interleukin-4 secreting mesenchymal stromal cells as an ""on-demand"" drug delivery system to modulate inflammation. Cytotherapy 19:1025-1034
Gallo, Jiri; Raska, Milan; Kriegova, Eva et al. (2017) Inflammation and its resolution and the musculoskeletal system. J Orthop Translat 10:52-67
Gibon, Emmanuel; Lu, Laura Y; Nathan, Karthik et al. (2017) Inflammation, ageing, and bone regeneration. J Orthop Translat 10:28-35

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