Human Umbilical Cord Mesenchymal Stem Cell-induced Osterix, Bone Morphogenetic Protein-2, and Tartrate-resistant Acid Phosphatase Expression in Osteoporotic Mandibular Bone

Abstract

Objectives The aim of this study was to prove that human umbilical cord mesenchymal stem cell (hUCMSC) therapy conducted according to the mandibular osteoporotic model will increase Osterix (Osx) and bone morphogenetic protein-2 (BMP-2) expression, while reducing tartrate-resistant acid phosphatase (TRAP) expression. PKH26 labeling proves that mandibular bone regeneration is produced by hUCMSCs induction.

Materials and Methods This study incorporated a true posttest only control group design. Twenty-five female Wistar rats were randomly divided into five groups consisting of the sham surgery (N) group, osteoporotic groups injected with gelatin for 4 weeks (G4) and 8 weeks (G8), and osteoporotic groups injected with hUCMSC-gelatin for 4weeks (SC4) and 8 weeks (SC8). All subjects were provided for BMP-2, Osx, and TRAP on immunohistochemistry examination and PKH-26 labeling.

Statistical Analysis All data were analyzed using ANOVA and Tukey HSD tests with p < 0.05 being considered as statistically significant.

Results Compared with other groups, the highest level of BMP-2 and Osx occurred in the sham surgery (N) and osteoporotic groups injected with hUCMSCs-gelatin (SC), while the lowest level of TRAP was found in SC4. During 4- and 8-week observation periods, the PKH 26 appeared green (fluorescent).

Conclusions hUCMSC demonstrates high-osteogenic activity and increased osteoporotic mandibular bone regeneration, as shown by increased expression of Osx and BMP-2 and decreased TRAP expression. From the labeling, PKH-26 proved that viable hUCMSCs in gelatin solvent can be present in the mandibular bone and be capable of promoting osteogenic differentiation and increasing mineralization and bone formation in the osteoporotic mandibular bone.

Keywords

stem cells - osteoporosis - bone - BMP 2 - osterix - TRAP - PKH-26

Publikationsverlauf

Artikel online veröffentlicht:
08. September 2020

• References

• 1 Bandela V, Munagapati B, Karnati RK, Venkata GRS, Nidudhur SR. Osteoporosis: its prosthodontic considerations - a review. J Clin Diagn Res 2015; 9 (12) ZE01-ZE04
• 2 Alkhader M, Aldawoodyeh A, Abdo N. Usefulness of measuring bone density of mandibular condyle in patients at risk of osteoporosis: a cone beam computed tomography study. Eur J Dent 2018; 12 (03) 363-368
• 3 Abraham A, Pullishery F. The effect of menopause on the periodontium - a review. IOSR J Dent Med Sci Ver I 2015; 14 (04) 79-82
• 4 Elani HW, Starr JR, Da Silva JD, Gallucci GO. Trends in dental implant use in the U.S., 1999 – 2016, and Projections to 2026. J Dent Res 2018; 97 (13) 1424-1430
• 5 Parithimarkalaignan S, Padmanabhan TV. Osseointegration: an update. J Indian Prosthodont Soc 2013; 13 (01) 2-6
• 6 Durão SF, Gomes PS, Silva-Marques JM. et al. Bone regeneration in osteoporotic conditions: healing of subcritical-size calvarial defects in the ovariectomized rat. Int J Oral Maxillofac Implants 2012; 27 (06) 1400-1408
• 7 Han YF, Tao R, Sun TJ, Chai JK, Xu G, Liu J. Optimization of human umbilical cord mesenchymal stem cell isolation and culture methods. Cytotechnology 2013; 65 (05) 819-827
• 8 Hendrijantini N, Kusumaningsih T, Rostiny R, Mulawardhana P, Danudiningrat CP, Rantam FA. A potential therapy of human umbilical cord mesenchymal stem cells for bone regeneration on osteoporotic mandibular bone. Eur J Dent 2018; 12 (03) 358-362
• 9 Hendrijantini N, Hartono P, Ari MDA, Rantan FA. Human umbilical cord mesenchymal stem-cell therapy to increase the density of osteoporotic mandibular bone. Eur J Dent 2019; 13 (01) 58-63
• 10 Chen PM, Lin CH, Li NT. et al. c-Maf regulates pluripotency genes, proliferation/self-renewal, and lineage commitment in ROS-mediated senescence of human mesenchymal stem cells. Oncotarget 2015; 6 (34) 35404-35418
• 11 Zhang C, Yuchi H, Sun L, Zhou X, Lin J. Human amnion-derived mesenchymal stem cells protect against UVA irradiation-induced human dermal fibroblast senescence, in vitro. Mol Med Rep 2017; 16 (02) 2016-2022
• 12 Hayman AR. Tartrate-resistant acid phosphatase (TRAP) and the osteoclast/immune cell dichotomy. Autoimmunity 2008; 41 (03) 218-223
• 13 Khajuria DK, Razdan R, Mahapatra DR. Description of a new method of ovariectomy in female rats. Rev Bras Reumatol 2012; 52 (03) 462-470
• 14 Kelp A, Abruzzese T, Wohrle S, Frajs V, Aicher WK. Labeling mesenchymal stromal cells with pkh26 or vibrant dii significantly diminishes their migration, but does not affect their viability, attachment, proliferation and differentiation capacities. J Tissue Sci Eng 2017; 8 (02) 199
• 15 Kitajima Y, Doi H, Ono Y. et al. Estrogen deficiency heterogeneously affects tissue specific stem cells in mice. Sci Rep 2015; 5 (12861) 12861
• 16 Sözen T, Özışık L, Başaran NÇ. An overview and management of osteoporosis. Eur J Rheumatol 2017; 4 (01) 46-56
• 17 Liu DB, Sui C, Wu TT, Wu LZ, Zhu YY, Ren ZH. Association of bone morphogenetic protein (BMP)/Smad signaling pathway with fracture healing and osteogenic ability in senile osteoporotic fracture in humans and rats. Med Sci Monit 2018; 24: 4363-4371
• 18 Mohamad N, Nabih ES, Zakaria ZM, Nagaty MM, Metwaly RG. Insight into the possible role of miR-214 in primary osteoporosis via osterix. J Cell Biochem 2019; 120 (09) 15518-15526
• 19 Rogers MB, Shah TA, Shaikh NN. Turning bone morphogenetic protein 2 (BMP 2) on and off in mesenchymal cells. J Cell Biochem 2015; 116 (10) 2127-2138
• 20 Sinha KM, Zhou X. Genetic and molecular control of osterix in skeletal formation. J Cell Biochem 2013; 114 (05) 975-984
• 21 Tu Q, Valverde P, Li S, Zhang J, Yang P, Chen J. Osterix overexpression in mesenchymal stem cells stimulates healing of critical-sized defects in murine calvarial bone. Tissue Eng 2007; 13 (10) 2431-2440
• 22 Li M, Chen X, Yan J. et al. Inhibition of osteoclastogenesis by stem cell-derived extracellular matrix through modulation of intracellular reactive oxygen species. Acta Biomater 2018; 71: 118-131
• 23 Winkler T, von P Roth, Matziolis G, Mehta M, Perka C, Duda GN. Dose-response relationship of mesenchymal stem cell transplantation and functional regeneration after severe skeletal muscle injury in rats. Tissue Eng Part A 2009; 15 (03) 487-492
• 24 Wang J, Rao S, Chu J. et al. A protein interaction network for pluripotency of embryonic stem cells. Nature 2006; 444 (7117) 364-368
• 25 Antebi B, Pelled G, Gazit D. Stem cell therapy for osteoporosis. Curr Osteoporos Rep 2014; 12 (01) 41-47