Effect of bone marrow stromal cells transferred with sonic hedgehog gene on osteoporosis in ovariectomized rats

doi:10.3877/cma.j.issn.2096-0263.2019.04.006

Abstract

Abstract:

Objective

To research the effect of bone marrow stromal cells transferred with sonic hedgehog gene (Shh-BMSCs) transplantation about osteoporosis in ovariectomized rats.

Methods

Forty female SD rats were divided into control group, model group, bone marrow stem cells (BMSCs) transplantation group and bone marrow stromal cells transferred with sonic hedgehog gene (Shh-MSCs) transplantation group, each group has ten animals. Rats in control group were only removed adipose tissue around the ovaries, not their ovaries. The rest of the three groups were performed bilateral ovariectomy, and osteoporosis model was established. Two months after modeling, rats in the BMSCs or Shh-BMSCs transplantation group were respectively given BMSCs or Shh-BM SCs via the rat tail intravenous (fourth generation, about 2*10⁶ stem cells), and the same amount of PBS was injected in the model group. Two weeks after treatment, levels of serum calcium, phosphorus, alkaline phosphatase were detected in rats, and bone mineral density was measured using dual energy X-ray, at the same timetibial form metrology was analyzed by micro -CT scan.

Results

(1) Compared with the control group, the bone trabecular thickness, trabecular bone quantity and bone volume fraction were significantly decreased in the model group (P<0.05). There was a significant reduction in the bone mineral density of the rat femur, lumbar spine and whole body as well as the levels of serum calcium and alkaline phosphatase, however the phosphorus was increased significantly (P<0.05). These findings indicate the successful preparation of the ostesporosis rat model. (2) Compared with the ovariedomized group, the levels of serum calcium and alkaline phosphatase were significantly higher in the BMSCs group, however the phosphorus was decreased significantly (P<0.05). Compared with the ovariedomized and BMSCs groups, the levels of serum calcium and alkaline phosphatase were significantly higher in the Shh-BMSCs group, however the phosphorus was decreased significantly (P<0.05). (3) Compared with the ovariedomized group, the bone mineral density of the rat femur, lumbar spine or whole body was significantly higher in the BMSCs group(P<0.05). Compared with the ovariedomized and BMSCs groups, the bone mineral density of the rat femur, lumbar spine or whole body was significantly higher in the Shh-BMSCs group(P<0.05). (4) Compared with the ovariedomized group, the number and thickness of trabeculae and bone volume fraclio was significantly higher in the BMSCs group(P<0.05). Compared with the ovariedomized and BMSCs groups, the number and thickness of trabeculae and bone volume fraclio was significantly higher in the Shh-BMSCs group(P<0.05).

Conclusion

Shh-BMSCs exerts therapeutic effects on osteoporosis in ovariectomized rats.

Key words: Marrow stromal cells, Sonic hedgehog protein, rat, Osteoporosis, Cell transplantaion

Yijia Jia, Chunshan Luo, Tingsheng Lu, Shudan Yao, Xingwei Pu, Jianwen Yang, Qiling Chen. Effect of bone marrow stromal cells transferred with sonic hedgehog gene on osteoporosis in ovariectomized rats[J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2019, 05(04): 211-216.

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Figures/Tables 4

References 21

19	Blank U, Karlsson G, Karlsson S. Signaling pathways governing stem-cell fate [J]. Blood, 2008, 111(2): 249-260.
20	Stucki U, Schmid J, Hammerle CF, et al. Temporal and local appearance of alkaline phosphatase activity in early stages of guided bone regeneration [J]. Clin Oral Implants Res, 2001, 12(2):121-127.
21	Peng Z, Tuukkanen J, Väänänen HK. Exercise can provide protection against bone loss and prevent the decrease in mechanical strength of femoral neck in ovariectomized rats [J]. J Bone Miner Res, 1994, 9(10): 1559-1564.
1	刘忠厚.骨质疏松学[M].北京:科学出版社, 1998: 529.
2	Honig S. Osteoporosis - new treatments and updates [J]. Bull NYU Hosp Jt Dis, 2010, 68(3): 166-170.
3	Papanicolaou DA, Wilder RL, Manolagas SC, et al. The pathophysiologic roles of interleukin-6 in human disease [J]. Ann Intern Med, 1998, 128(2): 127-137.
4	罗南萍,刘恒国.骨质疏松症与相关细胞因子的研究[J].放射免疫学杂志, 2007, 20(2): 132-136.
5	盛柩,谢培凤,王新祥.原发性骨质疏松症从脾论治及相关机制探讨[J].中华中医药杂志, 2012, 27(7): 1922-1926.
6	Vural P, Akgul C, Canbaz M. Effects of hormone replacement therapy on plasma pro-inflammatory and anti-inflammatory cytokines and some bone turnover markers in postmenopausal women [J]. Pharmacol Res, 2006, 54(4): 298-302.
7	Uemura H, Kamada M, Maegawa M, et al. Effect of hormone replacement therapy on the production of bone-resorbing cytokines by peripheral blood cells in postmenopausal women [J]. Horm Metab Res, 2005, 37(4): 226-230.
8	Eriksson PS, Perfilieva E, Björk-Eriksson T, et al. Neurogenesis in the adult human hippocampus [J]. Nat Med, 1998, 4(11): 1313-1317.
9	张晓玲,戴尅戎,汤亭亭,等.未分化骨髓间充质干细胞的免疫学特性研究[J].中华实验外科杂志, 2006, 23(2): 135-137.
10	Ma XL, Zhang XE, Jia YF, et al. Dexamethasone induces osteogenesis via regulation of hedgehog signalling molecules in rat mesenchymal stem cells [J]. Int Orthop, 2013, 37(7): 1399-1404.
11	Zhang XS, Linkhart TA, Chen ST, et al. Local ex vivo gene therapy with bone marrow stromal cells expressing human BMP4 promotes endosteal bone formation in mice [J]. J Gene Med, 2004, 6(1): 4-15.
12	Zimmermann S, Voss M, Kaiser S, et al. Lack of telomerase activity in human mesenchymal stem cells [J]. Leukemia, 2003, 17(6): 1146-1149.
13	Simonsen JL, Rosada C, Serakinci N, et al. Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells [J]. Nat Biotechnol, 2002, 20(6): 592-596.
14	Zhao YM, Li JY, Lan JP, et al. Cell cycle dependent telomere regulation by telomerase in human bone marrow mesenchymal stem cells [J]. Biochem Biophys Res Commun, 2008, 369(4): 1114-1119.
15	Bunnell BA, Estes BT, Guilak F, et al. Differentiation of adipose stem cells [J]. Methods Mol Biol, 2008,456:155-171
16	张颖,张博,张治国,等.从肝脾肾三脏探讨绝经后骨质疏松症的发病机理[J].中国中医基础医学杂志, 2012, 18(1): 36-42.
17	李雪琳.中西医结合治疗绝经妇女骨质疏松症30例[J].实用中医药杂志, 2012, 28(4): 285-286.
18	Kimura H1, Ng JM, Curran T. Transient inhibitiion of the hedgehog pathway in young mice causes permanent defects in bone structure [J]. Cancer Cell, 2008, 13(3): 1-46.

组别	钙（mmol/L）	磷（mmol/L）	碱性磷酸酶（U/L）	腰椎	股骨	全身
假手术组	3.59±0.63	3.69±0.23	201.31±29.23^abc	187.21±12.32	257.29±19.73	137.29±8.82
模型组	2.52±0.75^a	5.26±0.86^a	225±31.54	97.32±10.35^a	198.29±23.96^a	100.29±9.75^a
BMSCs组	3.08±0.64^ab	4.30±0.42^ab	132.29±19.25^a	122.19±10.84^ab	211.18±16.75^ab	112.02±11.45^ab
Shh-BMSCs组	3.31±0.68^abc	3.98±0.29^abc	170.52±25.06^ab	143.89±10.53^abc	230.62±22.51^abc	121.31±10.34^abc

组别	钙（mmol/L）	磷（mmol/L）	碱性磷酸酶（U/L）	腰椎	股骨	全身
假手术组	3.59±0.63	3.69±0.23	201.31±29.23^abc	187.21±12.32	257.29±19.73	137.29±8.82
模型组	2.52±0.75^a	5.26±0.86^a	225±31.54	97.32±10.35^a	198.29±23.96^a	100.29±9.75^a
BMSCs组	3.08±0.64^ab	4.30±0.42^ab	132.29±19.25^a	122.19±10.84^ab	211.18±16.75^ab	112.02±11.45^ab
Shh-BMSCs组	3.31±0.68^abc	3.98±0.29^abc	170.52±25.06^ab	143.89±10.53^abc	230.62±22.51^abc	121.31±10.34^abc

组别	例数（只）	骨小梁厚度（mm）	骨小梁数量（mm^-1）	骨小梁分离度（mm）	骨体积分数
假手术组	10	0.047±0.008	4.6729±0.131	0.208±0.036	0.196±0.021
模型组	10	0.025±0.009^a	3.127±0.263^a	0.369±0.531^a	0.098±0.009^a
BMSCs组	10	0.036±0.012^ab	3.988±0.284^ab	0.273±0.352^ab	0.136±0.021^ab
Shh-BMSCs组	10	0.042±0.010^abc	4.239±0.327^abc	0.239±0.326^abc	0.158±0.019^abc

组别	例数（只）	骨小梁厚度（mm）	骨小梁数量（mm^-1）	骨小梁分离度（mm）	骨体积分数
假手术组	10	0.047±0.008	4.6729±0.131	0.208±0.036	0.196±0.021
模型组	10	0.025±0.009^a	3.127±0.263^a	0.369±0.531^a	0.098±0.009^a
BMSCs组	10	0.036±0.012^ab	3.988±0.284^ab	0.273±0.352^ab	0.136±0.021^ab
Shh-BMSCs组	10	0.042±0.010^abc	4.239±0.327^abc	0.239±0.326^abc	0.158±0.019^abc

[1]	The Joint Surgery Branch of the Chinese Orthopedic Association, Guangdong’s Medical Association of Osteoporosis And Bone Mineral Research, Foshan Shunde 3rd Peoele’s Hospital. Clinical intervention guideline for using of anti-osteoporosis drugs in postoperative fragile hip fractures in China (2023 Edition) [J]. Chinese Journal of Joint Surgery(Electronic Edition), 2023, 17(06): 751-764.
[2]	Yan Wang, Jianxiong Ma, Shuang Lang, Benchao Dong, Aixian Tian, Yan Li, Lei Sun, Hongzhen Jin, Bin Lu, Ying Wang, Haohao Bai, Xinlong Ma. Research progress in application of exosomes in diagnosis and treatment of osteoporosis [J]. Chinese Journal of Joint Surgery(Electronic Edition), 2023, 17(05): 673-678.
[3]	Luqiao Pu, Yongqing Xu, Baochuang Qi, Hongxin Shi, Wei Lin, Pengfei Bu, Yan Bai, Zhifang Tang, Chuan Li. Prospectus on clinical intervention guideline for anti-osteoporosis drugs in postoperative fragile hip fractures in China (2023 edition) [J]. Chinese Journal of Joint Surgery(Electronic Edition), 2023, 17(05): 747-750.
[4]	Changjiao Sun, Peng Yu, Xu Cai, Yongwei Pan. Progress of shoulder arthroplasty in patients with osteoporosis [J]. Chinese Journal of Joint Surgery(Electronic Edition), 2023, 17(02): 253-260.
[5]	Joint Surgery Group of Chinese Orthopaedic Association., Shanghai Changzheng Hospital, Shanghai 200003, China. Management standard of chronic bone disease for community hospitals [J]. Chinese Journal of Joint Surgery(Electronic Edition), 2023, 17(01): 1-10.
[6]	Yixian Lu, Zhentao Zhang, Demeng Xia, Jialin Wang. Research progress of the regulating role and mechanism of macrophage polarization in osteoporosis [J]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2023, 18(06): 538-541.
[7]	Yueqi Chen, Rui Luo, Han Xiang, Yongyan Yu, Ting Yu. The causal association between osteoporosis and periodontitis: A two-sample Mendelian randomization study [J]. Chinese Journal of Stomatological Research(Electronic Edition), 2023, 17(04): 292-298.
[8]	Ranran Liu, Qianqian Fang, Zewen Tang. Effects of peripheral nerve injury on the proliferation and osteogenesis differentiation of bone marrow mesenchymal stem cells [J]. Chinese Journal of Neurotraumatic Surgery(Electronic Edition), 2023, 09(01): 7-11.
[9]	Juan Wang, Jun Gao, Yilan Zhou, Xiaohong Li, Bingwei Shi, Meizhen Pan. Relationship between serum IL-2, IL-17 and bone mineral density and their predictive value for osteoporosis [J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2023, 09(05): 295-300.
[10]	Xi Zhang, YeQing Liu, Xueying Kang, Bingbing Sun, Yan Liu, Liye Hu, Yaru Zhou. Correlation between serum ferritin and postmenopausal osteoporosis [J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2023, 09(03): 166-171.
[11]	Hang Xu, Yutao Cui, Guangkai Ren, He Liu, Yanbing Wang, Chuangang Peng, Dankai Wu. Screening and Bioinformatics Analysis of Core Genes of Osteoporosis [J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2023, 09(01): 18-22.
[12]	Chengyu Qin, Haonan Zhou, Yuanming Chen. The difference of anti-osteoporosis effect of Puerarin on different parts of bone in postmenopausal osteoporosis rats [J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2023, 09(01): 23-27.
[13]	Yan Wang, Wenjing Li, Hongzhi Lyu, Jing Li, Juan Wang, Chuan Ren, Yingze Zhang. Establishment and Evaluation of Clinical Predictive Model for Osteoporotic fracture [J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2022, 08(06): 350-360.
[14]	Yicheng Guo, Zhipeng Xie, Wanli Liu, Yang Zhao, Yansheng Che, Min Zhao. Characteristic analysis of precordial referred pain caused by osteoporotic vertebral compression fracture in the 12th thoracic vertebrae misdiagnosed as coronary heart disease [J]. Chinese Journal of Diagnostics(Electronic Edition), 2023, 11(02): 136-139.
[15]	Xiaohui Bai, Long Zhang, Yongfeng Wang, Yi Feng, Bin Zhao, Zhi Lyu, Chaojian Xu. Comparison of therapeutic effects between unilateral and bilateral percutaneous vertebroplasty for Kummell's disease [J]. Chinese Journal of Geriatrics Research(Electronic Edition), 2023, 10(02): 14-18.

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