Arne Mehrkens1, Ajay Matta2, Muhammad Zia Karim2, Sarah Kim2, Michael G Fehlings3, Stefan Schaeren4, William Mark Erwin5. 1. Krembil Research Institute, Toronto Western Hospital, 60 Leonard Ave, Toronto, Ontario M5T 2S8, Canada; Spine Surgery, University Hospital Basel, Spitalstr. 21, CH-4031 Basel, Switzerland. 2. Krembil Research Institute, Toronto Western Hospital, 60 Leonard Ave, Toronto, Ontario M5T 2S8, Canada. 3. Krembil Research Institute, Toronto Western Hospital, 60 Leonard Ave, Toronto, Ontario M5T 2S8, Canada; Division of Neurosurgery and Spine Program, University of Toronto, Toronto Western Hospital, 399 Bathurst St, Toronto, Ontario M5T 2S8, Canada. 4. Spine Surgery, University Hospital Basel, Spitalstr. 21, CH-4031 Basel, Switzerland. 5. Krembil Research Institute, Toronto Western Hospital, 60 Leonard Ave, Toronto, Ontario M5T 2S8, Canada; Division of Neurosurgery and Spine Program, University of Toronto, Toronto Western Hospital, 399 Bathurst St, Toronto, Ontario M5T 2S8, Canada; Division of Orthopaedic Surgery, University of Toronto, Toronto Western Hospital, 399 Bathurst St, Toronto, Ontario M5T 2S8, Canada; Division of Research, Canadian Memorial Chiropractic College, Toronto, Ontario M2H 3J1, Canada. Electronic address: mark.erwin@utoronto.ca.
Abstract
BACKGROUND CONTEXT: Degenerative disc disease (DDD) remains without an effective therapy and presents a costly burden to society. PURPOSE: Based upon prior reports concerning the effects of notochordal cell-conditioned medium (NCCM) on disc cells, we performed a proof of principle study to determine whether NCCM could reduce cytotoxic stress-induced apoptosis in human disc nucleus pulposus (NP) cells. STUDY DESIGN/ SETTING: This is an "in vitro" fundamental or basic science study. METHODS: Nucleus pulpous cells derived from 15 patients undergoing spinal surgery were treated with interleukin (IL)-1β and Fas ligand or etoposide in the presence of NCCM. We determined pro- or antiapoptotic events using activated caspase assays and determined genomic regulation of apoptosis using polymerase chain reaction arrays validated using Western blotting methods. We interrogated cellular apoptotic regulation using JC-1 dye and flow cytometry and performed enzyme-linked immunosorbent assays to evaluate NP inflammatory cytokine secretion. RESULTS: Notochordal cell-conditioned medium inhibits cytotoxic stress-induced caspase-9 and -3/7 activities and maintains the mitochondrial membrane potential in human NP cells, thereby suppressing the intrinsic apoptotic pathway. Gene expression analysis revealed the X-linked inhibitor of apoptosis protein as a key player responsible for evading etoposide-induced apoptosis in the presence of NCCM, and we verified these data using Western blotting. Enzyme-linked immunosorbent assay results revealed distinct differences in IL-6 and IL-8 secretions by NP cells in response to etoposide in the presence of NCCM. CONCLUSIONS: Here we demonstrate for the first time that NCCM reduces cytotoxic stress-induced apoptosis in human NP cells. Soluble factors present in NCCM could be harnessed for the development of novel therapeutics for the treatment of DDD.
BACKGROUND CONTEXT: Degenerative disc disease (DDD) remains without an effective therapy and presents a costly burden to society. PURPOSE: Based upon prior reports concerning the effects of notochordal cell-conditioned medium (NCCM) on disc cells, we performed a proof of principle study to determine whether NCCM could reduce cytotoxic stress-induced apoptosis in human disc nucleus pulposus (NP) cells. STUDY DESIGN/ SETTING: This is an "in vitro" fundamental or basic science study. METHODS: Nucleus pulpous cells derived from 15 patients undergoing spinal surgery were treated with interleukin (IL)-1β and Fas ligand or etoposide in the presence of NCCM. We determined pro- or antiapoptotic events using activated caspase assays and determined genomic regulation of apoptosis using polymerase chain reaction arrays validated using Western blotting methods. We interrogated cellular apoptotic regulation using JC-1 dye and flow cytometry and performed enzyme-linked immunosorbent assays to evaluate NP inflammatory cytokine secretion. RESULTS: Notochordal cell-conditioned medium inhibits cytotoxic stress-induced caspase-9 and -3/7 activities and maintains the mitochondrial membrane potential in human NP cells, thereby suppressing the intrinsic apoptotic pathway. Gene expression analysis revealed the X-linked inhibitor of apoptosis protein as a key player responsible for evading etoposide-induced apoptosis in the presence of NCCM, and we verified these data using Western blotting. Enzyme-linked immunosorbent assay results revealed distinct differences in IL-6 and IL-8 secretions by NP cells in response to etoposide in the presence of NCCM. CONCLUSIONS: Here we demonstrate for the first time that NCCM reduces cytotoxic stress-induced apoptosis in human NP cells. Soluble factors present in NCCM could be harnessed for the development of novel therapeutics for the treatment of DDD.
Authors: Ruhang Tang; Liufang Jing; Vincent P Willard; Chia-Lung Wu; Farshid Guilak; Jun Chen; Lori A Setton Journal: Stem Cell Res Ther Date: 2018-03-09 Impact factor: 6.832
Authors: Weihang Li; Shilei Zhang; Dong Wang; Huan Zhang; Quan Shi; Yuyuan Zhang; Mo Wang; Ziyi Ding; Songjie Xu; Bo Gao; Ming Yan Journal: Front Cell Dev Biol Date: 2022-02-10
Authors: Frances C Bach; Deepani W Poramba-Liyanage; Frank M Riemers; Jerome Guicheux; Anne Camus; James C Iatridis; Danny Chan; Keita Ito; Christine L Le Maitre; Marianna A Tryfonidou Journal: Front Cell Dev Biol Date: 2022-03-14