Non-invasive in vivo molecular imaging of intra-articularly transplanted immortalized bone marrow stem cells for osteoarthritis treatment

Bou-Yue Peng; Chi-Sheng Chiou; Navneet Kumar Dubey; Sung-Hsun Yu; Yue-Hua Deng; Feng-Chou Tsai; Han-Sun Chiang; Ying-Hua Shieh; Wei-Hong Chen; Win-Ping Deng

doi:10.18632/oncotarget.21315

Non-invasive in vivo molecular imaging of intra-articularly transplanted immortalized bone marrow stem cells for osteoarthritis treatment

Oncotarget. 2017 Sep 27;8(57):97153-97164. doi: 10.18632/oncotarget.21315. eCollection 2017 Nov 14.

Authors

Bou-Yue Peng¹, Chi-Sheng Chiou^{2

3}, Navneet Kumar Dubey^{4

5}, Sung-Hsun Yu^{4

6}, Yue-Hua Deng^{4

7}, Feng-Chou Tsai⁸, Han-Sun Chiang⁷, Ying-Hua Shieh^{9

10}, Wei-Hong Chen⁴, Win-Ping Deng^{2

4

11}

Affiliations

¹ Oral and Maxillofacial Surgery Section, Department of Dentistry, Taipei Medical University Hospital, Taipei, Taiwan.
² School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan.
³ Division of Allergy, Immunology and Rheumatology, Department of Internal Medicine, Taipei Medical University Hospital, Taipei, Taiwan.
⁴ Stem Cell Research Center, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan.
⁵ Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan.
⁶ Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan.
⁷ Department of Life Science, Fu Jen Catholic University, Taipei, Taiwan.
⁸ Department of Stem Cell Research, Cosmetic Clinic Group, Taipei, Taiwan.
⁹ Department of Family Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
¹⁰ Department of Family Medicine, Taipei Medical University Hospital, Taipei, Taiwan.
¹¹ Graduate Institute of Basic Medicine, Fu Jen Catholic University, Taipei, Taiwan.

Abstract

Pathophysiology of osteoarthritis (OA) is characterized by progressive loss of articular cartilage in the knee-joints. To impart regenerative ability in lowly metabolizing chondrocytes, the bone marrow stem cells (BMSCs) has recently been recognized as a superior alternative treatment for OA. However, study of primary BMSCs-mediated chondrogenesis is difficult due to progressive cellular aging and replicative senescence. To obtain a therapeutic cell population for OA, BMSCs were immortalized by human papilloma virus (HPV)-16 E6/E7 along with mCherry luciferase (mCL), a gene marker for non-invasive imaging, and designated as iBMSCs-mCL. Next, their cell morphology, population doubling time (PDT) and colony forming ability (CFU) were evaluated. Furthermore, pluripotency and immunophenotypic markers were investigated. To deduce therapeutic ability, iBMSCs-mCL were intra-articularly injected into right knee of anterior cruciate ligament transaction (ACLT)-OA mice model and tracked through non-invasive bioluminescence imaging. Cell morphology of iBMSCs-mCL was similar to parental BMSCs. PDT and CFU ability of iBMSCs-mCLs were significantly increased. Pluripotency and immunophenotypic markers were highly expressed in iBMSC-mCL. Long-term survival and tri-lineage differentiation particularly chondrogenic potential of iBMSCs-mCL were also demonstrated in vitro and then in vivo which was monitored through non-invasive imaging. Intensive bioluminescent signals in iBMSCs-mCL administered knee-joint indicated a marked in vivo survival and proliferation of iBMSCs-mCL. Immunohistochemical staining for type II collagen (IHC of Col II) and alcian blue & safranin o staining of proteoglycans also corroborated cartilage regeneration by iBMSCs-mCL. Conclusively, iBMSCs-mCL maintains stemness and in vivo cartilage regeneration potential suggesting a promising avenue for development of OA therapeutics.

Keywords: cartilage regeneration; human papilloma virus (HPV)-16 E6/E7; immortalized bone marrow stem cells (iBMSCs); molecular imaging; osteoarthritis (OA).