Irradiation alters the differentiation potential of bone marrow mesenchymal stem cells

Wang,Yu; Zhu,Guoying; Wang,Jianping; Chen,Junxiang

doi:10.3892/mmr.2015.4539

Irradiation alters the differentiation potential of bone marrow mesenchymal stem cells

Authors:
- Yu Wang
- Guoying Zhu
- Jianping Wang
- Junxiang Chen
View Affiliations

Affiliations: Department of Radiation Protection, Institute of Radiation Medicine, Fudan University, Shanghai 200032, P.R. China
Published online on: November 10, 2015 https://doi.org/10.3892/mmr.2015.4539
Pages: 213-223
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Bone injury following radiotherapy has been confirmed by epidemiological and animal studies. However, the underlying mechanism remains to be elucidated and no preventive or curative solution has been identified for this bone loss. The present study aimed to investigate the irradiation‑altered osteogenesis and adipogenesis of bone marrow mesenchymal stem cells (BMSCs). BMSCs were derived and exposed to γ‑irradiation at doses of 0, 0.25, 0.5, 1, 2, 5 and 10 Gy. Cell viability was assessed using a 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5 diphenyl tetrazolium bromide assay, and clonal expansion in vitro was detected by colony forming unit assessment. The osteogenic differentiation ability was demonstrated by alkaline phosphatase (ALP) activity, ALP staining and mineralization alizarin red staining, and the adipogenic differentiation ability was determined using Oil O red staining. The osteogenesis‑associated genes, RUNX2, ALP, osteocalcin (OCN) and adipogenesis‑associated genes, PPAR‑γ and C/EBPα, were detected using reverse transcription‑quantitative polymerase chain reaction analyses. The protein expression levels of RUNX2, ALP and PPAR‑γ were detected using western blotting. Compared with the control, significant decreases in the proliferation, ALP activity and mineralization ability of the BMSCs were observed in the γ‑irradiation group, with a high level of correlation with the exposure dose. However, no significant changes were observed in the area of Oil red O positive staining. The mRNA levels of RUNX2, ALP and OCN were decreased (P<0.05), however, no significant changes were observed in the levels of C/EBPα and PPAR‑γ. The protein expression levels of RUNX2 and ALP were decreased in the irradiated BMSCs, however, no significant difference was observed in the protein expression of PPAR‑γ. Irradiation inhibited the osteogenic and adipogenic ability of the BMSCs, and the osteogenic differentiation was decreased. The results of the present study provided evidence to assist in further elucidating radiotherapy‑associated side effects on the skeleton.

View Figures

View References

1	Jeremic B: Radiation therapy. Hematol Oncol Clin North Am. 18:1–12. 2004. View Article : Google Scholar : PubMed/NCBI
2	Darby S, McGale P, Correa C, Taylor C, Arriagada R, Clarke M, Cutter D, Davies C, Ewertz M, Godwin J, et al Early Breast Cancer Trialists' Collaborative Group (EBCTCG): Effect of radiotherapy after breast-conserving surgery on 10-year recurrence and 15-year breast cancer death: Meta-analysis of individual patient data for 10,801 women in 17 randomised trials. Lancet. 378:1707–1716. 2011. View Article : Google Scholar : PubMed/NCBI
3	Cancer survivorship - United States, 1971–2001. MMWR Morb Mortal Wkly Rep. 53:526–529. 2004.
4	Dinshaw KA, Budrukkar AN, Chinoy RF, Sarin R, Badwe R, Hawaldar R and Shrivastava SK: Profile of prognostic factors in 1022 Indian women with early-stage breast cancer treated with breast-conserving therapy. Int J Radiat Oncol Biol Phys. 63:1132–1141. 2005. View Article : Google Scholar : PubMed/NCBI
5	Agrawal S: Late effects of cancer treatment in breast cancer survivors. South Asian J Cancer. 3:112–115. 2014. View Article : Google Scholar : PubMed/NCBI
6	Williams HJ and Davies AM: The effect of X-rays on bone: A pictorial review. Eur Radiol. 16:619–633. 2006. View Article : Google Scholar
7	Baxter NN, Habermann EB, Tepper JE, Durham SB and Virnig BA: Risk of pelvic fractures in older women following pelvic irradiation. JAMA. 294:2587–2593. 2005. View Article : Google Scholar : PubMed/NCBI
8	Green DE and Rubin CT: Consequences of irradiation on bone and marrow phenotypes, and its relation to disruption of hematopoietic precursors. Bone. 63:87–94. 2014. View Article : Google Scholar : PubMed/NCBI
9	Coquard R: Late effects of ionizing radiations on the bone marrow. Cancer Radiother. 1:792–800. 1997. View Article : Google Scholar
10	Jia D, Gaddy D, Suva LJ and Corry PM: Rapid loss of bone mass and strength in mice after abdominal irradiation. Radiat Res. 176:624–635. 2011. View Article : Google Scholar : PubMed/NCBI
11	Rodríguez JP, Astudillo P, Ríos S and Pino AM: Involvement of adipogenic potential of human bone marrow mesenchymal stem cells (MSCs) in osteoporosis. Curr Stem Cell Res Ther. 3:208–218. 2008. View Article : Google Scholar : PubMed/NCBI
12	Asano S: Current status of hematopoietic stem cell transplantation for acute radiation syndromes. Int J Hematol. 95:227–231. 2012. View Article : Google Scholar : PubMed/NCBI
13	Shao L, Luo Y and Zhou D: Hematopoietic stem cell injury induced by ionizing radiation. Antioxid Redox Signal. 20:1447–1462. 2014. View Article : Google Scholar :
14	Christensen DM, Iddins CJ and Sugarman SL: Ionizing radiation injuries and illnesses. Emerg Med Clin North Am. 32:245–265. 2014. View Article : Google Scholar
15	Heylmann D, Rödel F, Kindler T and Kaina B: Radiation sensitivity of human and murine peripheral blood lymphocytes, stem and progenitor cells. Biochim Biophys Acta. 1846.121–129. 2014.
16	Friedenstein A and Kuralesova AI: Osteogenic precursor cells of bone marrow in radiation chimeras. Transplantation. 12:99–108. 1971. View Article : Google Scholar : PubMed/NCBI
17	Zhang L, Peng LP, Wu N and Li LP: Development of bone marrow mesenchymal stem cell culture in vitro. Chin Med J (Engl). 125:1650–1655. 2012.
18	Bidwell JP, Alvarez MB, Hood MJ and Childress P: Functional impairment of bone formation in the pathogenesis of osteoporosis: The bone marrow regenerative competence. Curr Osteoporos Rep. 11:117–125. 2013. View Article : Google Scholar : PubMed/NCBI
19	Bethel M, Chitteti BR, Srour EF and Kacena MA: The changing balance between osteoblastogenesis and adipogenesis in aging and its impact on hematopoiesis. Curr Osteoporos Rep. 11:99–106. 2013. View Article : Google Scholar : PubMed/NCBI
20	James AW, Shen J, Khadarian K, Pang S, Chung G, Goyal R, Asatrian G, Velasco O, Kim J, Zhang X, et al: Lentiviral delivery of PPARγ shRNA alters the balance of osteogenesis and adipogenesis, improving bone microarchitecture. Tissue Eng Part A. 20:2699–2710. 2014. View Article : Google Scholar : PubMed/NCBI
21	Justesen J, Stenderup K, Ebbesen EN, Mosekilde L, Steiniche T and Kassem M: Adipocyte tissue volume in bone marrow is increased with aging and in patients with osteoporosis. Biogerontology. 2:165–171. 2001. View Article : Google Scholar : PubMed/NCBI
22	Post S, Abdallah BM, Bentzon JF and Kassem M: Demonstration of the presence of independent pre-osteoblastic and pre-adipocytic cell populations in bone marrow-derived mesenchymal stem cells. Bone. 43:32–39. 2008. View Article : Google Scholar : PubMed/NCBI
23	Gimble JM, Zvonic S, Floyd ZE, Kassem M and Nuttall ME: Playing with bone and fat. J Cell Biochem. 98:251–266. 2006. View Article : Google Scholar : PubMed/NCBI
24	Nicolay NH, Lopez PR, Debus J and Huber PE: Mesenchymal stem cells - A new hope for radiotherapy-induced tissue damage? Cancer Lett. 366:133–140. 2015. View Article : Google Scholar : PubMed/NCBI
25	Soleimani M and Nadri S: A protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow. Nat Protoc. 4:102–106. 2009. View Article : Google Scholar : PubMed/NCBI
26	Qiu J, Zhu G, Chen X, Shao C and Gu S: Combined effects of γ-irradiation and cadmium exposures on osteoblasts in vitro. Environ Toxicol Pharmacol. 33:149–157. 2012. View Article : Google Scholar : PubMed/NCBI
27	Chen X, Zhu G, Gu S, Jin T and Shao C: Effects of cadmium on osteoblasts and osteoclasts in vitro. Environ Toxicol Pharmacol. 28:232–236. 2009. View Article : Google Scholar : PubMed/NCBI
28	Siclari VA, Zhu J, Akiyama K, Liu F, Zhang X, Chandra A, Nah HD, Shi S and Qin L: Mesenchymal progenitors residing close to the bone surface are functionally distinct from those in the central bone marrow. Bone. 53:575–586. 2013. View Article : Google Scholar : PubMed/NCBI
29	Wernle JD, Damron TA, Allen MJ and Mann KA: Local irradiation alters bone morphology and increases bone fragility in a mouse model. J Biomech. 43:2738–2746. 2010. View Article : Google Scholar : PubMed/NCBI
30	Turner RT, Iwaniec UT, Wong CP, Lindenmaier LB, Wagner LA, Branscum AJ, Menn SA, Taylor J, Zhang Y, Wu H, et al: Acute exposure to high dose γ-radiation results in transient activation of bone lining cells. Bone. 57:164–173. 2013. View Article : Google Scholar : PubMed/NCBI
31	Tewarie RDSN, Hurtado A, Grotenhuis JA and Oudega M: Bone marrow stromal cell survival, migration, and differentiation following acute and delayed transplantation into the moderately contused adult rat thoracic spinal cord. Cell Res. 18:S1002008. View Article : Google Scholar
32	Chen Q, Shou P, Zhang L, Xu C, Zheng C, Han Y, Li W, Huang Y, Zhang X, Shao C, et al: An osteopontin-integrin interaction plays a critical role in directing adipogenesis and osteogenesis by mesenchymal stem cells. Stem Cells. 32:327–337. 2014. View Article : Google Scholar :
33	Cao X, Wu X, Frassica D, Yu B, Pang L, Xian L, Wan M, Lei W, Armour M, Tryggestad E, et al: Irradiation induces bone injury by damaging bone marrow microenvironment for stem cells. Proc Natl Acad Sci USA. 108:1609–1614. 2011. View Article : Google Scholar : PubMed/NCBI
34	Gimble JM, Zvonic S, Floyd ZE, Kassem M and Nuttall ME: Playing with bone and fat. J Cell Biochem. 98:251–266. 2006. View Article : Google Scholar : PubMed/NCBI
35	Abdallah BM and Kassem M: New factors controlling the balance between osteoblastogenesis and adipogenesis. Bone. 50:540–545. 2012. View Article : Google Scholar
36	Agata H, Yamazaki M, Uehara M, Hori A, Sumita Y, Tojo A and Kagami H: Characteristic differences among osteogenic cell populations of rat bone marrow stromal cells isolated from untreated, hemolyzed or Ficoll-treated marrow. Cytotherapy. 14:791–801. 2012. View Article : Google Scholar : PubMed/NCBI
37	Hendry JH: The cellular basis of long-term marrow injury after irradiation. Radiother Oncol. 3:331–338. 1985. View Article : Google Scholar : PubMed/NCBI
38	Ashkenazi A and Dixit VM: Apoptosis control by death and decoy receptors. Curr Opin Cell Biol. 11:255–260. 1999. View Article : Google Scholar : PubMed/NCBI
39	Deng Y, Wu S, Zhou H, Bi X, Wang Y, Hu Y, Gu P and Fan X: Effects of a miR-31, Runx2, and Satb2 regulatory loop on the osteogenic differentiation of bone mesenchymal stem cells. Stem Cells Dev. 22:2278–2286. 2013. View Article : Google Scholar : PubMed/NCBI
40	Li J, Zhang N, Huang X, Xu J, Fernandes JC, Dai K and Zhang X: Dexamethasone shifts bone marrow stromal cells from osteoblasts to adipocytes by C/EBPalpha promoter methylation. Cell Death Dis. 4:e8322013. View Article : Google Scholar : PubMed/NCBI
41	Viccica G, Francucci CM and Marcocci C: The role of PPARγ for the osteoblastic differentiation. J Endocrinol Invest. 33(Suppl): 9–12. 2010.
42	Justesen J, Stenderup K, Eriksen EF and Kassem M: Maintenance of osteoblastic and adipocytic differentiation potential with age and osteoporosis in human marrow stromal cell cultures. Calcif Tissue Int. 71:36–44. 2002. View Article : Google Scholar : PubMed/NCBI