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Would three BMPs at low concentration be better than one at high concentration? An experimental study with rat osteoprogenitor cells

Stanislaw Moskalewski1, Anna Hyc1, Anna Osiecka-Iwan1
Pubmed: 38842074

Abstract

Background: Bone morphogenetic proteins (BMPs) are used in clinical practice for stimulation of bone formation, but often evoke serious complications. Recent studies demonstrated that BMPs involved in early stages of bone formation are species specific. In cattle dominate BMP7, growth differentiation factor 5 (GDF5) and NEL-like protein 1 (NELL1) while in rats BMP2, BMP5 and BMP6. The purpose of the study was to compare the action of the species specific BMPs on the osteoprogenitor cells. Thus, rat osteoprogenitor cells were exposed to one BMP in a high dose and three of them at 1/3 of the former.

Materials and methods: Isolated rat osteoprogenitor cells were treated in culture with different concentrations of BMP2, BMP5 and BMP6 or with lower concentration of combinations of these cytokines. Activity of alkaline phosphatase, calcium deposition and mRNA level for transcription factor SP7 (osterix) and tissue non-specific alkaline phosphatase (TNAP) served as indicators of BMPs effect.

Results: BMPs stimulated all studied parameters in comparison with control cultures, but no statistically significant differences were observed between the action of a large dose of one cytokine and a combination of cytokines given at lower concentrations.

Conclusions: Three BMPs used in a low dose exert similar effect as the one used at high dose. Since the BMPs stimulate different receptors and activate different signaling pathways the use of the mixture of properly chosen BMPs at low concentration may give better results than the single one at high concentration and may avoid untoward effects.

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References

  1. Abuna RPF, Oliveira FS, Ramos JIR, et al. Selection of reference genes for quantitative real-time polymerase chain reaction studies in rat osteoblasts. J Cell Physiol. 2018; 234(1): 749–756.
  2. Aoki H, Fujii M, Imamura T, et al. Synergistic effects of different bone morphogenetic protein type I receptors on alkaline phosphatase induction. J Cell Sci. 2001; 114(Pt 8): 1483–1489.
  3. Asahina I, Sampath TK, Nishimura I, et al. Human osteogenic protein-1 induces both chondroblastic and osteoblastic differentiation of osteoprogenitor cells derived from newborn rat calvaria. J Cell Biol. 1993; 123(4): 921–933.
  4. Bonor J, Adams EL, Bragdon B, et al. Initiation of BMP2 signaling in domains on the plasma membrane. J Cell Physiol. 2012; 227(7): 2880–2888.
  5. Celeste AJ, Iannazzi JA, Taylor RC, et al. Identification of transforming growth factor beta family members present in bone-inductive protein purified from bovine bone. Proc Natl Acad Sci U S A. 1990; 87(24): 9843–9847.
  6. Cheng H, Jiang W, Phillips FM, et al. Osteogenic activity of the fourteen types of human bone morphogenetic proteins (BMPs). J Bone Joint Surg Am. 2003; 85(8): 1544–1552.
  7. Courvoisier A, Sailhan F, Laffenêtre O, et al. French Study Group of BMP in Orthopedic Surgery. Bone morphogenetic protein and orthopaedic surgery: can we legitimate its off-label use? Int Orthop. 2014; 38(12): 2601–2605.
  8. da Silva Madaleno C, Jatzlau J, Knaus P. BMP signalling in a mechanical context — implications for bone biology. Bone. 2020; 137: 115416.
  9. Ducy P, Karsenty G. The family of bone morphogenetic proteins. Kidney Int. 2000; 57(6): 2207–2214.
  10. Ebara S, Nakayama K. Mechanism for the action of bone morphogenetic proteins and regulation of their activity. Spine (Phila Pa 1976). 2002; 27(16 Suppl 1): S10–S15.
  11. Ebisawa T, Tada K, Kitajima I, et al. Characterization of bone morphogenetic protein-6 signaling pathways in osteoblast differentiation. J Cell Sci. 1999; 112 ( Pt 20): 3519–3527.
  12. Eggerschwiler B, Canepa DD, Pape HC, et al. Automated digital image quantification of histological staining for the analysis of the trilineage differentiation potential of mesenchymal stem cells. Stem Cell Res Ther. 2019; 10(1): 69.
  13. Florencio-Silva R, Sasso GR, Sasso-Cerri E, et al. Biology of bone tissue: structure, function, and factors that influence bone cells. Biomed Res Int. 2015; 2015: 421746.
  14. Friedman MS, Long MW, Hankenson KD. Osteogenic differentiation of human mesenchymal stem cells is regulated by bone morphogenetic protein-6. J Cell Biochem. 2006; 98(3): 538–554.
  15. Gregory CA, Gunn WG, Peister A, et al. An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. Anal Biochem. 2004; 329(1): 77–84.
  16. Halloran D, Durbano HW, Nohe A. Bone morphogenetic protein-2 in development and bone homeostasis. J Dev Biol. 2020; 8(3).
  17. Haubruck P, Tanner MC, Vlachopoulos W, et al. Comparison of the clinical effectiveness of bone morphogenic protein (BMP) -2 and -7 in the adjunct treatment of lower limb nonunions. Orthop Traumatol Surg Res. 2018; 104(8): 1241–1248.
  18. Hefley T, Cushing J, Brand JS. Enzymatic isolation of cells from bone: cytotoxic enzymes of bacterial collagenase. Am J Physiol. 1981; 240(5): C234–C238.
  19. Hojo H, Ohba S. Sp7 action in the skeleton: its mode of action, functions, and relevance to skeletal diseases. Int J Mol Sci. 2022; 23(10).
  20. Hustedt JW, Blizzard DJ. The controversy surrounding bone morphogenetic proteins in the spine: a review of current research. Yale J Biol Med. 2014; 87(4): 549–561.
  21. Hyc A, Moskalewski S, Osiecka-Iwan A. Growth factors in the initial stage of bone formation, analysis of their expression in chondrocytes from epiphyseal cartilage of rat costochondral junction. Folia Histochem Cytobiol. 2021; 59(3): 178–186.
  22. Hyc A, Osiecka-Iwan A, Moskalewski S. Could BMPs Therapy Be Improved if BMPs Were Used in Composition Acting during Bone Formation in Endochondral Ossification? Int J Mol Sci. 2022; 23(18).
  23. Idaszek J, Jaroszewicz J, Choińska E, et al. Toward osteomimetic formation of calcium phosphate coatings with carbonated hydroxyapatite. Biomater Adv. 2023; 149: 213403.
  24. Iwan A, Moskalewski S, Hyc A. Growth factor profile in calcified cartilage from the metaphysis of a calf costochondral junction, the site of initial bone formation. Biomed Rep. 2021; 14(6): 54.
  25. James AW, LaChaud G, Shen J, et al. A review of the clinical side effects of bone morphogenetic protein-2. Tissue Eng Part B Rev. 2016; 22(4): 284–297.
  26. James AW, Shen J, Tsuei R, et al. NELL-1 induces Sca-1+ mesenchymal progenitor cell expansion in models of bone maintenance and repair. JCI Insight. 2017; 2(12).
  27. Kanjilal D, Cottrell JA. Bone morphogenetic proteins (BMPs) and bone regeneration. Methods Mol Biol. 2019; 1891: 235–245.
  28. Katagiri T, Watabe T. Bone Morphogenetic Proteins. Cold Spring Harb Perspect Biol. 2016; 8(6).
  29. Krishnakumar GS, Roffi A, Reale D, et al. Clinical application of bone morphogenetic proteins for bone healing: a systematic review. Int Orthop. 2017; 41(6): 1073–1083.
  30. Lavery K, Swain P, Falb D, et al. BMP-2/4 and BMP-6/7 differentially utilize cell surface receptors to induce osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells. J Biol Chem. 2008; 283(30): 20948–20958.
  31. Li Bo, Wang H, Qiu G, et al. Synergistic effects of vascular endothelial growth factor on bone morphogenetic proteins induced bone formation in vivo: influencing factors and future research directions. Biomed Res Int. 2016; 2016: 2869572.
  32. Liu L, Chen Y, Song D, et al. BMP9 is a potential therapeutic agent for use in oral and maxillofacial bone tissue engineering. Biochem Soc Trans. 2020; 48(3): 1269–1285.
  33. Liu M, Goldman G, MacDougall M, et al. BMP signaling pathway in dentin development and diseases. Cells. 2022; 11(14).
  34. Lowery JW, Rosen V. Bone morphogenetic protein-based therapeutic approaches. Cold Spring Harb Perspect Biol. 2018; 10(4).
  35. Luu HH, Song WX, Luo X, et al. Distinct roles of bone morphogenetic proteins in osteogenic differentiation of mesenchymal stem cells. J Orthop Res. 2007; 25(5): 665–677.
  36. May RD, Frauchiger DA, Albers CE, et al. Application of cytokines of the bone morphogenetic protein (BMP) family in spinal fusion — effects on the bone, intervertebral disc and mesenchymal stromal cells. Curr Stem Cell Res Ther. 2019; 14(8): 618–643.
  37. Miyazono K, Maeda S, Imamura T. BMP receptor signaling: transcriptional targets, regulation of signals, and signaling cross-talk. Cytokine Growth Factor Rev. 2005; 16(3): 251–263.
  38. Moon JS, Kim SD, Ko HM, et al. Twist1 suppresses cementoblast differentiation. Dent J (Basel). 2018; 6(4).
  39. Nakashima K, Zhou X, Kunkel G, et al. The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell. 2002; 108(1): 17–29.
  40. Narasimhulu CA, Singla DK. BMP-7 attenuates sarcopenia and adverse muscle remodeling in diabetic mice via alleviation of lipids, inflammation, HMGB1, and pyroptosis. Antioxidants (Basel). 2023; 12(2).
  41. Nickel J, Mueller TD. Specification of BMP signaling. Cells. 2019; 8(12).
  42. Peng H, Usas A, Olshanski A, et al. VEGF improves, whereas sFlt1 inhibits, BMP2-induced bone formation and bone healing through modulation of angiogenesis. J Bone Miner Res. 2005; 20(11): 2017–2027.
  43. Pignatti E, Zeller R, Zuniga A, et al. Smad4 is required to induce digit ray primordia and to initiate the aggregation and differentiation of chondrogenic progenitors in mouse limb buds. Development. 2012; 139(22): 4250–4260.
  44. Salazar VS, Gamer LW, Rosen V. BMP signalling in skeletal development, disease and repair. Nat Rev Endocrinol. 2016; 12(4): 203–221.
  45. Sampath TK, Maliakal JC, Hauschka PV, et al. Recombinant human osteogenic protein-1 (hOP-1) induces new bone formation in vivo with a specific activity comparable with natural bovine osteogenic protein and stimulates osteoblast proliferation and differentiation in vitro. J Biol Chem. 1992; 267(28): 20352–20362.
  46. Sánchez-Duffhues G, Hiepen C, Knaus P, et al. Bone morphogenetic protein signaling in bone homeostasis. Bone. 2015; 80: 43–59.
  47. Simic P, Culej JB, Orlic I, et al. Systemically administered bone morphogenetic protein-6 restores bone in aged ovariectomized rats by increasing bone formation and suppressing bone resorption. J Biol Chem. 2006; 281(35): 25509–25521.
  48. Takemoto RC, Fajardo M, Kirsch T, et al. Quantitative assessment of the bone morphogenetic protein expression from alternate bone graft harvesting sites. J Orthop Trauma. 2010; 24(9): 564–566.
  49. Tanjaya J, Zhang Y, Lee S, et al. Efficacy of intraperitoneal administration of pegylated NELL-1 for bone formation. Biores Open Access. 2016; 5(1): 159–170.
  50. Dijke Pt, Yamashita H, Sampath TK, et al. Identification of type I receptors for osteogenic protein-1 and bone morphogenetic protein-4. J Biol Chem. 1994; 269(25): 16985–16988.
  51. Termaat MF, Den Boer FC, Bakker FC, et al. Bone morphogenetic proteins. Development and clinical efficacy in the treatment of fractures and bone defects. J Bone Joint Surg Am. 2005; 87(6): 1367–1378.
  52. Vijayan V, Gupta S, Gupta S. Bone morphogenetic protein-5, a key molecule that mediates differentiation in MC3T3E1 osteoblast cell line. Biofactors. 2017; 43(4): 558–566.
  53. Vimalraj S. Alkaline phosphatase: structure, expression and its function in bone mineralization. Gene. 2020; 754: 144855.
  54. Vukicevic S, Grgurevic L. BMP-6 and mesenchymal stem cell differentiation. Cytokine Growth Factor Rev. 2009; 20(5-6): 441–448.
  55. Wang CKL, Omi M, Ferrari D, et al. Function of BMPs in the apical ectoderm of the developing mouse limb. Dev Biol. 2004; 269(1): 109–122.
  56. Wozney JM. The bone morphogenetic protein family and osteogenesis. Mol Reprod Dev. 1992; 32(2): 160–167.
  57. Wozney JM. Overview of bone morphogenetic proteins. Spine (Phila Pa 1976). 2002; 27(16 Suppl 1): S2–S8.
  58. Wutzl A, Rauner M, Seemann R, et al. Bone morphogenetic proteins 2, 5, and 6 in combination stimulate osteoblasts but not osteoclasts in vitro. J Orthop Res. 2010; 28(11): 1431–1439.
  59. Yagiela JA, Woodbury DM. Enzymatic isolation of osteoblasts from fetal rat calvaria. Anat Rec. 1977; 188(3): 287–306.
  60. Yu PB, Beppu H, Kawai N, et al. Bone morphogenetic protein (BMP) type II receptor deletion reveals BMP ligand-specific gain of signaling in pulmonary artery smooth muscle cells. J Biol Chem. 2005; 280(26): 24443–24450.
  61. Zhang L, Luo Q, Shu Yi, et al. Transcriptomic landscape regulated by the 14 types of bone morphogenetic proteins (BMPs) in lineage commitment and differentiation of mesenchymal stem cells (MSCs). Genes Dis. 2019; 6(3): 258–275.
  62. Zhang X, Zara J, Siu RK, et al. The role of NELL-1, a growth factor associated with craniosynostosis, in promoting bone regeneration. J Dent Res. 2010; 89(9): 865–878.
  63. Zhao W, He B, Zhou Ao, et al. D-RADA16-RGD-Reinforced nano-hydroxyapatite/polyamide 66 ternary biomaterial for bone formation. Tissue Eng Regen Med. 2019; 16(2): 177–189.
  64. Zhu L, Liu Y, Wang Ao, et al. Application of BMP in bone tissue engineering. Front Bioeng Biotechnol. 2022; 10: 810880.
  65. Zuzarte-Luís V, Montero JA, Rodriguez-León J, et al. A new role for BMP5 during limb development acting through the synergic activation of Smad and MAPK pathways. Dev Biol. 2004; 272(1): 39–52.