Stem-cell therapy for ovariectomy-induced osteoporosis in rats: a comparison of three treatment modalities

Stem Cells and Cloning: Advances and Applications Dovepress open access to scientific and medical research Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 88.198.20.149 on 10-Aug-2019 For personal use only. Open Access Full Text Article Stem-cell therapy for ovariectomy-induced osteoporosis in rats: a comparison of three treatment modalities This article was published in the following Dove Press journal: Stem Cells and Cloning: Advances and Applications Mir Sadat-Ali 1 Dakheel A Al-Dakheel 1 Sulaiman A AlMousa 1 Fawaz M AlAnii 1 Waleed Y Ebrahim 1 Hussain K AlOmar 1 Hasan N AlSayed 1 Sadananda Acharya 2 Hussain AlHawaj 3 1 Department of Orthopaedic Surgery, College of Public Health, 3Institute of Research and Medical Consultations, Imam Abdul Rahman Bin Faisal University, Dammam, Saudi Arabia 2 Background: Recent studies have shown that ovariectomy-induced osteoporosis in rats can be reversed by infusion of osteoblasts cultured from mesenchymal stem cells (MSCs). This study compares the influence of MSCs, osteoblasts, and exosomes derived from osteoblasts for the treatment of osteoporosis. Methods: Osteoporosis was induced in 40 female Sprague Dawley rats by performing ovariectomy. After 12 weeks, bone marrow was harvested and MSCs separated from bonemarrow aspirate as described by Piao et al. After 15 days, autologous osteogenically differentiated cells from the MSCs were available. Exosomes were isolated from osteoblasts by modification of the technique described by Ge et al. MSCs and osteoblasts (106 cells in 0.5 mL normal saline) and exosomes (100 µg protein) were injected into the tail veins of the animals. Animals were euthanized after 12 weeks and femurs and lumbar spines dissected and analyzed using high-resolution peripheral quantitative computed tomography. Results: When compared to the control group, osteoblast-treated animals showed significant differences in all parameters compared, with P-values ranging between <0.002 and <0.0001. Comparison among osteoblasts, MSCs, and exosomes, showed that osteoblasts had positive and statistically significant new-bone formation. The comparison for the spine was similar to the distal femur for osteoblasts. Conclusion: This study showed robust positive bone-forming changes after osteoblast injection in the distal femur and the spine when compared to controls, MSCs, and exosomes. Keywords: osteoporosis, ovariectomy, osteoblasts, mesenchymal stem cells (MSCs), exosomes Introduction Correspondence: Mir Sadat-Ali University Department of Orthopaedic Surgery, POBOX 40071, AlKhobar 31952, Saudi Arabia Tel +966 050 584 8281 Fax +96 603 882 0887 Email Osteoporosis is a chronic and debilitating disease of aging. Postmenopausal osteoporosis has come close to becoming an epidemic in the developed and developing world, with increased morbidity and mortality. The “silent disease", as it is rightfully termed, presents with a fracture due to minimal trauma its first indication.5 The true cost of managing osteoporosis-related fractures in Saudi Arabia is still unknown, but a recent study indicated that by 2050, the lifetime cost of treating osteoporosis-related femoral fractures in Saudi Arabia may reach over SR35 billion annually (US$9.33 billion).6 The cost of managing osteoporosis is increasing exponentially, but new modalities of treatment are lagging behind. We still are dependent on antiresorptives, such as bisphosphonates, anabolic agents like synthetic parathyroid hormone, and the newer 17 submit your manuscript | www.dovepress.com Stem Cells and Cloning: Advances and Applications 2019:12 17–25 DovePress © 2019 Sadat-Ali et al. This work is published and licensed by Dove Medical Press Limited. 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Sadat-Ali et al monoclonal antibody denosumab, and all have their limitations and complications of use.7–12 Preclinical studies have shown promising results in the management of ovariectomy-induced osteoporosis. Kiernan et al2 showed that mesenchymal stem cells (MSCs) injected into a mouse model led to improved bone formation and reversed microarchitecture, suggesting that MSC injections may be used to combat osteoporosis, whereas Sadat-Ali et al1 reported that culture-expanded osteoblasts, when injected in ovariectomized (Ovx) rats, were effective in significantly increasing bone formation. Exosomes, which are bioactive microvesicles, are secreted by MSCs, and osteoblasts are nanoparticles of 30–100 nm in size and carry proteins and RNAs.13 MSCderived exosomes have been found to mediate in promoting healing of tissue and repair of acute and chronic injuries.14 With the increasing number of aged people around the world and increase in the longevity of the human race, osteoporosis is bound to trouble health-care economics, added to the fact that research and development of new osteoporosis drugs are at a standstill. Stem-cell therapy opens a new avenue in the treatment of osteoporosis if the reversal of osteoporosis in animal studies is confirmed. The objective of this study was to compare the efficacy of culture expanded MSCs, osteoblasts, and osteoblast-derived exosomes on the effect of Ovx-induced osteoporosis in rats. Methods Ethical approval was obtained from Imam AbdulRahman Bin Faisal University, Dammam, Saudi Arabia (vide number 2015116/2017), 40 Sprague Dawley female rats were used as our model to study reversal of osteoporosis. The rats were housed and handled in accordance with the National Advisory Committee for Laboratory Animal Research guidelines. Animals were accommodated with total mobility, fed a standard diet, and the room maintained at 26°C. Ovariectomy was done at 1 month of age to cause osteoporosis. At 12 weeks, a bone biopsy was performed to look at the quality of bone. Bone marrow was aspirated at the time of the biopsy. From the bone-marrow aspirate of the individual rats, using the technique described by Piao et al,3 MSCs were separated. In this study, no MSCs from other species, such as human or BALB/c mouse were used. For each individual rat, MSCs obtained from the bonemarrow aspirate and osteoblasts cultured from the cell 18 Powered by TCPDF (www.tcpdf.org) submit (...truncated)