Synthetic PTHrP1-36 and its role in promoting alveolar bone formation

doi:10.19439/j.sjos.2020.06.004

Abstract

Abstract: PURPOSE: To synthetise mouse parathyroid hormone related protein 1-36 (PTHrP1-36) and to study its role in promoting alveolar bone formation. METHODS: pGEX-2TK/ PTHrP1-36 was constructed and PTHrP1-36 was expressed. The role of PTHrP1-36 in promoting alveolar bone formation was observed by treating bone marrow cells in vitro and ovariectomy-induced maxillofacial osteoporosis in vivo. Statistical analysis was performed using SPSS 10.0 software package. RESULTS: The results of PCR analysis showed that pGEX-2TK/ PTHrP1-36 plasmid was correct templates for expressing PTHrP1-36. The molecular weights obtained by SDS-PAGE were the same as the theoretical value after IPTG induction. Scale up production was carried out by 3-hour-induction with 1mM IPTG which was the optimal induction factors. Bone marrow cell culture revealed that synthetic PTHrP1-36 increased the number of total fibroblastic colony forming units (CFU-f), ALP positive CFU-f and calcific nodules. After PTHrP1-36 was subcutaneously administered to ovariectomy mice, density and volume of alveolar bone were improved, as showed by X-ray and histochemistry staining of collagen-l and H-E staining. CONCLUSIONS: Synthetic PTHrP1-36 can promote bone marrow cell osteogenic differentiation and increase alveolar bone formation.

Key words: Parathyroid hormone related protein, Alveolar bone, Osteoprogenitor, Synthetic protein

CLC Number:

R782.1

LI Qin, LYU Chun-hua, CHEN Lin, LIU Xian, WANG Hua, SUN Wen. Synthetic PTHrP1-36 and its role in promoting alveolar bone formation[J]. Shanghai Journal of Stomatology, 2020, 29(6): 580-585.

References

[1] 关晓月, 钱华, 边专. 卵泡刺激素在女性绝经后骨质疏松及口腔骨吸收疾病中的研究[J]. 口腔医学研究, 2017, 33(5): 574-576.
[2] 文艺, 杨鸿旭, 刘倩, 等. 骨髓间充质干细胞联合血管内皮祖细胞修复骨质疏松性牙槽骨缺损[J]. 中国组织工程研究, 2016, 20(19): 2748-2755.
[3] 李晶, 艾红军, 洪岩松. 骨质疏松症SD大鼠废用残根对周围牙槽骨组织影响的研究[J].中国医科大学学报, 2015, 44(5): 400-404.
[4] Persson GR, Berglund J, Persson RE, et al.Prediction of hip and hand fractures in older persons with or without a diagnosis of periodontitis[J]. Bone, 2011, 48(3): 552-556.
[5] Suresh S, Kumar TS, Saraswathy PK, et al.Periodontitis and bone mineral density among pre and post menopausal women: a comparative study[J]. J Indian Soc Periodontol, 2010, 14(1): 30-34.
[6] Dempster DW, Cosman F, Kurland ES, et al.Effects of daily treatment with parathyroid hormone on bone microarchitecture and turnover in patients with osteoporosis: a paired biopsy study[J]. J Bone Miner Res, 2001, 16(10): 1846-1853.
[7] Zhang J, Liao L, Li Y, et al.Parathyroid hormone-related peptide (1-34) promotes tooth eruption and inhibits osteogenesis of dental follicle cells during tooth development[J]. J Cell Physiol, 2019, 234(7): 11900-11911.
[8] Wang CJ, McCauley LK. Osteoporosis and periodontitis[J]. Curr Osteoporos Rep, 2016, 14(6): 284-291.
[9] Binte Anwar R, Tanaka M, Kohno S, et,al. Relationship between porotic changes in alveolar bone and spinal osteoporosis[J]. J Dent Res, 2007, 86(1): 52-57.
[10] Luna-Cabrera F, Justicia-Rull EA, Caricol-Pérez MP, et al.Incidence of hypercalcemia, hypercalciuria and related factors in patients treated with recombinant human parathyroid hormone (1-84)[J]. Minerva Med, 2012, 103(2): 103-110.
[11] Liu M, Kurimoto P, Zhang J, et al.Sclerostin and DKK1 inhibition preserves and augments alveolar bone volume and architecture in rats with alveolar bone loss[J]. J Dental Res, 2018, 97(9) : 1031-1038.
[12] Sun W, Wu J, Huang L, et al.PTHrP nuclear localization and carboxyl terminus sequences modulate dental and mandibular development in part via the action of p27[J]. Endocrinology, 2016, 157(4): 1372-1384.
[13] Miao D, Tong XK, Chan GK, et al.Parathyroid hormone-related peptide stimulates osteogenic cell proliferation through protein kinase C activation of the Ras/mitogen-activated protein kinase signaling pathway[J]. J Biol Chem, 2001, 276(34): 32204-32213.
[14] Miao D, He B, Jiang Y, et al.Osteoblast-derived PTHrP is a potent endogenous bone anabolic agent that modifies the therapeutic efficacy of administered PTH 1-34[J]. J Clin Invest, 2005, 115(9): 2402-2411.
[15] Xue Y, Karaplis AC, Hendy GN, et al.Genetic models show that parathyroid hormone and 1,25-dihydroxyvitamin D3 play distinct and synergistic roles in postnatal mineral ion homeostasis and skeletal development[J]. Hum Mol Genet, 2005, 14(11): 1515-1528.