Co-overexpression of OPN, IGF-1 and CNTF augment the therapeutic effect of DPSC on spinal cord injury

Ning Tao; Xiwen Dong; Chao Liu; Lin Lv; Fanqi Hu; Huan Zhang; Xue Li; Panpan Geng; Han Duan; Chu-Tse Wu; Hua Wang

doi:10.1016/j.reth.2023.11.004

Co-overexpression of OPN, IGF-1 and CNTF augment the therapeutic effect of DPSC on spinal cord injury

Regen Ther. 2023 Nov 17:24:651-661. doi: 10.1016/j.reth.2023.11.004. eCollection 2023 Dec.

Authors

Ning Tao¹, Xiwen Dong², Chao Liu³, Lin Lv⁴, Fanqi Hu⁵, Huan Zhang⁴, Xue Li^{1

4}, Panpan Geng⁴, Han Duan⁴, Chu-Tse Wu⁴, Hua Wang^{1

4}

Affiliations

¹ College of Life Science, Anhui Medical University, Hefei, 230032, PR China.
² Department of Dermatology, Air Force Medical Center, PLA, Beijing, 100142, PR China.
³ Department of Orthopaedics, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310009, PR China.
⁴ Beijing Institute of Radiation Medicine, Beijing, 100850, PR China.
⁵ Department of Orthopaedics, the Fourth Medical Centre, Chinese PLA General Hospital, Beijing, 100853, PR China.

Abstract

Background aims: Spinal cord injury (SCI) is one of the most complex and destructive diseases of the nervous system, which can lead to permanent loss of tactile perception. But existing treatment methods have limited effects. To establish a novel method that may be therapeutic in repairing the injured spinal cord, gene-modified dental pulp stem cells (DPSCs) were injected in situ.

Methods: Adenovirus carrying osteopontin (OPN), Insulin-like growth factor 1 (IGF-1) and cailiary-derived neurotrophic factor (CNTF) (Ad-OIC) was constructed. After modified with Ad-OIC, supernatant of DPSC were co-cultured with HT-22 cells and the effect of DPSC-OIC on the HT-22 cells were evaluated via Cell Counting Kit-8 (CCK-8) assay, Real-Time polymerase chain reaction (PCR) analysis, laser confocal microscopy and fluorescence activating cell sorter (FACS). DPSC-OIC were injected in the lesion area of injured spinal cord and the survival time of transplanted cells were measured by bioluminescence imaging system. The recovery of the injured spinal cord was evaluated by behavioral score, radiological evaluation and immunopathological analysis.

Results: DPSC-OIC could enhance the proliferation and axon growth of HT-22 cells, and protect HT-22 cells from H₂O₂ induced apoptosis. The transplanted DPSC-Null or DPSC-OIC could survive for more than two weeks in local injection site. DPSC-OIC treatment could increase Basso-Mouse Scale (BMS) scores, improve Magnetic Resonance Imaging (MRI) manifestation and promote bladder function recovery. Less apoptotic neurons and more proliferative cells were found in the lesion area of DPSC-OIC treated spinal cord. Nestin⁺ cells and neural stem cell marker (Sox2) were both up-regulated after DPSC-OIC treatment. Additionally, inhibitory extracellular matrix proteoglycan Neural/Glial Antigen 2 (NG2) was down-regulated and axon growth promotive factor fibronectin was up-regulated after both DPSC-Null (DPSCs infected with Ad-Null) and DPSC-OIC treatments.

Conclusions: DPSC-OIC could be a novel effective method for treating SCI.

Keywords: Dental pulp stem cells; Neurons; Regenerative medicine; Spinal cord injury.