CC BY 4.0 · Eur J Dent 2023; 17(04): 1106-1113
DOI: 10.1055/s-0042-1758786
Original Article

Osteoinductive and Osteogenic Capacity of Freeze-Dried Bovine Bone Compared to Deproteinized Bovine Bone Mineral Scaffold in Human Umbilical Cord Mesenchymal Stem Cell Culture: An In Vitro Study

Andreas Pratama Nugraha
1   Magister Program of Clinical Medicine, Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia
2   Department of Oral and Maxillofacial Surgery, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia
,
David B. Kamadjaja
2   Department of Oral and Maxillofacial Surgery, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia
,
Ni Putu Mira Sumarta
2   Department of Oral and Maxillofacial Surgery, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia
,
Andra Rizqiawan
2   Department of Oral and Maxillofacial Surgery, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia
,
Coen Pramono
2   Department of Oral and Maxillofacial Surgery, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia
,
Anita Yuliati
3   Departement of Dental Material, Faculty of Dental medicine Universitas Airlangga, Surabaya, Indonesia
,
Eryk Hendrianto
4   Stem Cell Research and Developmental Center, Universitas Airlangga, Surabaya, Indonesia
,
Mohammad Zeshaan Rahman
5   Department of Oral and Maxillofacial Surgery, Pioneer Dental College and Hospital, Dhaka, Bangladesh
› Institutsangaben

Abstract

Objective Freeze-dried bovine bone scaffold (FDBB) or decellularized FDBB (dc-FDBB) was developed as an ideal scaffold with osteoinductive properties. This research aims to compare the osteoinductive properties marked by the expression of runt-related transcription factor-2 (RUNX2) and Osterix (OSX) and the osteogenic capacity of these scaffolds imbued with human umbilical cord mesenchymal stem cells (hUCMSCs).

Materials and Methods This study was performed in five experimental groups: a negative control group (C-) of hUCMSCs with a normal growth medium, a positive control group (C + ) of hUCMSCs with an osteogenic medium, experimental group 1 (E1) with an FDBB conditioned medium (CM), and experimental group 2 (E2) with a dc-FDBB-CM, and a third experimental group (E3) consisting of a DBBM-CM. Alizarin red staining was performed to qualitatively assess osteoinductive capacity. RUNX2 and OSX expression was quantified using real-time quantification polymerase chain reaction with two replications on day six (D6) and day 12 (D12) as fold changes.

Results This experiment revealed that hUCMSCs were positively expressed by CD73, CD90, and CD105 but were not expressed by CD34. Alizarin red staining showed that E1 had the most calcium deposition on D6 and D12, followed by E3 and then E2 The RUNX2 and OSX expression was higher in E1 but this difference was not significant. The OSX expression in E1,E2,E3 was lower on D12 and C+ of OSX had the highest expression. There was a significant difference of fold change measured between all groups (p < 0.05), and there was no significant difference between any of the groups treated with OSX and RUNX2 on D6 and D12.

Conclusion FDBB osteoinduction and osteogenic capacity were higher when compared with DBBM and dc-FDBB.



Publikationsverlauf

Artikel online veröffentlicht:
04. Januar 2023

© 2023. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)

Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India

 
  • References

  • 1 Alfotawi R, Ayoub A. Reconstruction of maxillofacial bone defects: Contemporary methods and future techniques. Am J Adv Med Sci 2014; 2 (01) 18-27
  • 2 Rana M, Warraich R, Kokemüller H. et al. Reconstruction of mandibular defects - clinical retrospective research over a 10-year period. Head Neck Oncol 2011; 3 (01) 23
  • 3 Zhao R, Yang R, Cooper P, Khurshid Z, Shavandi A, Ratnayake J. Bone graft and substitute in dentistry: a review of trends and developments. Molecules (MDPI) 2021; 26 (03) 1-27
  • 4 Pereira HF, Cengiz F, Silva S, Reis L. Oliveira. Scaffold and coatings for bone regeneration. J Mat Sci Mat Med 2020; 31 (27) 1-16
  • 5 Zhang J, Xie C, Lu Y, Zhou M. Potential antigen involved in delayed xenograft rejection in a Ggta1/Cmah Dko pig to monkey model. Sci Rep 2017; 30 (02) 12-25
  • 6 Agrawal A, Mehrotra D, Mohammad S, Singh RK, Kumar S, Pal US. Randomized control trial of non-vascularized fibular and iliac crest graft for mandibular reconstruction. J Oral Biol Craniofac Res 2012; 2 (02) 90-96
  • 7 Mahyudin F, Utomo DN, Suroto H, Gaol IL. Immunogenicity of bone graft using xenograft freeze-dried cortical bovine, allograft freeze-dried cortical New Zealand white rabbit, xenograft hydroxyapatite bovine, and xenograft demineralized bone matrix bovine in bone defect of femoral diaphysis white rabbit experimental study in vivo. The Veterinary Medicine International Conference KnE Life Sciences. 2017: 344-355
  • 8 Kamadjaja DB, Satriyo H, Setyawan A. et al. Analyses of bone regeneration capacity of freeze dried bovine bone and combined deproteinized demineralized bovine bone particles in mandibular defects: the potential application of biological form of bovine bone filler. Eur J Dent 2021; 21: 11-23
  • 9 Ansari M. Bone tissue regeneration: biology, strategies and interface studies. Prog Biomater 2019; 8 (04) 223-237
  • 10 Kangwannarongkul T, Subbalekha K, Vivatbutsiri P, Suwanwela J. Gene expression and microcomputed tomography analysis of grafted bone using deproteinized bovine bone and freeze-dried human bone. Int J Oral Maxillofac Implants 2018; 33 (03) 541-548
  • 11 Matsuoka F, Takeuchi I, Agata H. et al. Morphology-based prediction of osteogenic differentiation potential of human mesenchymal stem cells. PLoS One 2013; 8 (02) e55082
  • 12 Nugraha AP, Narmada IB, Ernawati DS. et al. Osteogenic potential of gingival stromal progenitor cells cultured in platelet rich fibrin is predicted by core-binding factor subunit-α1/Sox9 expression ratio (in vitro). F1000 Res 2018; 7: 1134
  • 13 Lorenzo J, Horowitz M, Choi Y, Takayanagi H. Osteoimmunology. 1st ed. London: Elsevier; 2011: 100-140
  • 14 Hendrijatini N, Hartono C, Daniati R, Hong Maya R, Kuntjoro M, Ari MD. Human umbilical cord mesenchymal stem cell induced osterix, bone morphogenetic protein-2 and tartrate -resistant acid phospatase expression in osteoporotic mandibular bone. Eur J Dent 2021; 5: 84-89
  • 15 Hendrijantini N, Hartono P. Phenotype characteristics and osteogenic differentiation potential of human mesenchymal stem cells derived from amnion membrane (HAMSCs) and umbilical cord (HUC-MSCs). Acta Inform Med 2019; 27 (02) 72-77
  • 16 Filho GS, Caballé-Serrano J, Sawada K. et al. Conditioned medium of demineralized freeze-dried bone activates gene expression in periodontal fibroblasts in vitro. J Periodontol 2015; 86 (06) 827-834
  • 17 Zhong S, He X, Li Y, Lou X. Conditioned medium enhances osteogenic differentiation of induced pluripotent stem cell-derived mesenchymal stem cells. Tissue Eng Regen Med 2019; 16 (02) 141-150
  • 18 Rai R, Raval R, Khandeparker RVS, Chidrawar SK, Khan AA, Ganpat MS. Tissue engineering: step ahead in maxillofacial reconstruction. J Int Oral Health 2015; 7 (09) 138-142
  • 19 Kamadjaja DB. Purwati, Triakoso N. Bovine bone xenograft scaffold seeded with human umbilibal cord mesenchymal stem cell to reconstruct segmental defect in a dog's mandible: a preliminary study. Biochem Cell Arch 2019; 2 (04) 4871-4876
  • 20 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
  • 21 Zou L, Luo Y, Chen M. et al. A simple method for deriving functional MSCs and applied for osteogenesis in 3D scaffolds. Sci Rep 2013; 3: 2243
  • 22 Chen YS, Pelekanos RA, Ellis RL, Horne R, Wolvetang EJ, Fisk NM. Small molecule mesengenic induction of human induced pluripotent stem cells to generate mesenchymal stem/stromal cells. Stem Cells Transl Med 2012; 1 (02) 83-95
  • 23 Chen G, Deng C, Li YP. TGF-β and BMP signaling in osteoblast differentiation and bone formation. Int J Biol Sci 2012; 8 (02) 272-288
  • 24 Tirkkonen L, Haimi S, Huttunen S. et al. Osteogenic medium is superior to growth factors in differentiation of human adipose stem cells towards bone-forming cells in 3D culture. Eur Cell Mater 2013; 25: 144-158
  • 25 Chen X, Wang Z, Duan N, Zhu G, Schwarz EM, Xie C. Osteoblast-osteoclast interactions. Connect Tissue Res 2018; 59 (02) 99-107
  • 26 Hagh FM, Nourzina M, Mortazavi Y. Different methylation patterns of RUNX2, OSX, DLC5 and BSP in osteoblastic differentiation of mesenchymal stem cells. Cell J 2014; 17 (01) 71-82
  • 27 Furuya M, Shimono N, Okamoto M. Fabrication of biocomposites composed of natural rubber latex and bone tissue derived from MC3T3–E1 mouse preosteoblastic cells. Nanocomposites 2017; 3 (02) 1-17
  • 28 Marupanthorn K, Tantrawatpan C, Kheolamai P, Tantikanlayaporn D, Manochantr S. Bone morphogenetic protein-2 enhances the osteogenic differentiation capacity of mesenchymal stromal cells derived from human bone marrow and umbilical cord. Int J Mol Med 2017; 39 (03) 654-662
  • 29 Sinha KM, Zhou X. Genetic and molecular control of osterix in skeletal formation. J Cell Biochem 2013; 114 (05) 975-984
  • 30 Liu F, Akiyama Y, Tai S. et al. Changes in the expression of CD106, osteogenic genes, and transcription factors involved in the osteogenic differentiation of human bone marrow mesenchymal stem cells. J Bone Miner Metab 2008; 26 (04) 312-320
  • 31 Mukherjee A, Rotwein P. Akt promotes BMP2-mediated osteoblast differentiation and bone development. J Cell Sci 2009; 122 (Pt 5): 716-726
  • 32 Valenti MT, Dalle Carbonare L, Donatelli L, Bertoldo F, Zanatta M, Lo Cascio V. Gene expression analysis in osteoblastic differentiation from peripheral blood mesenchymal stem cells. Bone 2008; 43 (06) 1084-1092
  • 33 Tang W, Li Y, Osimiri L, Zhang C. Osteoblast-specific transcription factor Osterix (Osx) is an upstream regulator of Satb2 during bone formation. J Biol Chem 2011; 286 (38) 32995-33002
  • 34 He S, Yang S, Zhang Y, Li X. LncRNA ODIR1 inhibits osteogenic differentiation of hUC-MSCs through theFBXO25/H2BK120ub/H3K4me3/OSX axis. Cell Death Dis 2019; 4 (02) 1-18
  • 35 Graves T, Oates T, Garlet G. Osteoimmunology in the oral cavity, in osteoimmunology interaction of the immune and skeletal system. Philadelphia: Elsevier Inc; 2008: 325-344
  • 36 Merivaara A, Zini J, Koivunotko E. et al. Preservation of biomaterials and cells by freeze-drying: Change of paradigm. J Control Release 2021; 336 (10) 480-498
  • 37 Amirazad H, Dadashpour M, Zarghami N. Application of decellularized bone matrix as a bioscaffold in bone tissue engineering. J Biol Eng 2022; 16 (01) 1-15
  • 38 Woo JS, Fishbein MC, Reemtsen B. Histologic examination of decellularized porcine intestinal submucosa extracellular matrix (CorMatrix) in pediatric congenital heart surgery. Cardiovasc Pathol 2016; 25 (01) 12-17
  • 39 Nart J, Barallat L, Jimenez D. et al. Radiographic and histological evaluation of deproteinized bovine bone mineral vs. deproteinized bovine bone mineral with 10% collagen in ridge preservation. A randomized controlled clinical trial. Clin Oral Implants Res 2017; 28 (07) 840-848
  • 40 Barradas AM, Fernandes HA, Groen N. et al. A calcium-induced signaling cascade leading to osteogenic differentiation of human bone marrow-derived mesenchymal stromal cells. Biomaterials 2012; 33 (11) 3205-3215
  • 41 Guo X, Jiang H, Zong X. et al. The implication of the notch signaling pathway in biphasic calcium phosphate ceramic-induced ectopic bone formation: a preliminary experiment. J Biomed Mater Res A 2020; 108 (05) 1035-1044
  • 42 Chen QY, Liu SY, Liu AY. Biochemical and physical characterization of mesenchymal stromal cell along the time course of directed differentiation. Sci Rep 2016; 8 (02) 1-14
  • 43 National Measurement System. Good Practice guide for the application of Quantitative Polymerase Chain Reaction, 1st ed. 2013: 1-80