Journal of Biosciences

, 44:23 | Cite as

Mechanistic role of perfusion culture on bone regeneration

  • Bhaskar BirruEmail author
  • Naveen Kumar Mekala
  • Sreenivasa Rao Parcha


Bone tissue engineering (BTE) aims to develop engineered bone tissue to substitute conventional bone grafts. To achieve this, culturing the cells on the biocompatible three-dimensional (3D) scaffold is one alternative approach. The new functional bone tissue regeneration could be feasible by the synergetic combinations of cells, biomaterials and bioreactors. Although the field of biomaterial design/development for BTE applications attained reasonable success, development of suitable bioreactor remains still a major challenge. Tissue engineering bioreactors provide the microenvironment required for neo-tissue regeneration, and also can be used to study the physio-chemical cues effect on cell proliferation and differentiation in order to produce functional tissue. In this direction, various bioreactors have been developed and evaluated for the successful development of engineered bone tissue. Continues assessment of tissue development and limitations of the bioreactors lead to the progression of perfusion flow bioreactor system. Improvements in perfusion reactor system were able to yield multiple tissue engineered constructs with uniform cell distribution, easy to operate protocols and also effectively handled for the functional tissue development to meet the adequate supply of engineered graft for clinical application.


Bioreactor bone tissue engineering Physico-chemical cues perfusion reactor 3D scaffolds 



  1. Araujo JV, Cunha-Reis C, Rada T, et al. 2009 Dynamic culture of osteogenic cells in biomimetically coated poly(caprolactone) nanofibre mesh constructs. Tissue Eng. Part A, 16 557–563CrossRefGoogle Scholar
  2. Baharvand H, Hashemi SM and Shahsavani M 2008 Differentiation of human embryonic stem cells into functional hepatocyte-like cells in a serum-free adherent culture condition. Differentiation 76 465–477CrossRefGoogle Scholar
  3. Barron MJ, Goldman J, Tsai CJ and Donahue SW 2012 Perfusion flow enhances osteogenic gene expression and the infiltration of osteoblasts and endothelial cells into three-dimensional calcium phosphate scaffolds. Int. J. Biomater. 2012 17–20CrossRefGoogle Scholar
  4. Beşkardeş IG, Hayden RS, Glettig DL, Kaplan DL and Gümüşderelioğlu M 2017 Bone tissue engineering with scaffold-supported perfusion co-cultures of human stem cell-derived osteoblasts and cell line-derived osteoclasts. Process Biochem. 59 303–311CrossRefGoogle Scholar
  5. Bhaskar B, Mekala NK, Baadhe RR and Rao PS 2014 Role of signaling pathways in mesenchymal stem cell differentiation. Curr. Stem Cell Res. Ther. 9 508–512CrossRefGoogle Scholar
  6. Bhaskar B, Owen R, Bahmaee H, Rao PS and Reilly GC 2017 Design and assessment of a dynamic perfusion bioreactor for large bone tissue engineering scaffolds. Appl. Biochem. Biotechnol. 185 555–563CrossRefGoogle Scholar
  7. Bhaskar B, Owen R, Bahmaee H, Wally Z, Rao PS and Reilly GC 2018 Composite porous scaffold of Polyethylene glycol (PEG)/Polylactic acid (PLA) support improved bone matrix deposition in vitro compared to PLAonly scaffolds. J. Biomed. Mater. Res. Part A 106 1334–1340CrossRefGoogle Scholar
  8. Burdick JA and Vunjak-Novakovic G2009 engineered microenvironments for controlled stem cell differentiation. Tissue Eng. Part A 15 205–219Google Scholar
  9. Carmona-Moran CA and Wick TM 2015 transient growth factor stimulation improves chondrogenesis in static culture and under dynamic conditions in a novel shear and perfusion bioreactor. Cell. Mol. Bioeng. 8 267–277CrossRefGoogle Scholar
  10. Damania A, Kumar A, Sarin SK and Kumar A2017 Optimized performance of the integrated hepatic cell-loaded cryogel-based bioreactor with intermittent perfusion of acute liver failure plasma. J. Biomed. Mater. Res. Part B Appl. Biomater. 106 259–269Google Scholar
  11. Faghihi F, Papadimitropoulos A, Martin I and Eslaminejad MB 2014 Effect of purmorphamine on osteogenic differentiation of human mesenchymal stem cells in a three-dimensional dynamic culture system. Cell. Mol. Bioeng. 7 575–584CrossRefGoogle Scholar
  12. Filipowska J, Reilly GC and Osyczka AM 2016 A single short session of media perfusion induces osteogenesis in hBMSCs cultured in porous scaffolds, dependent on cell differentiation stage. Biotechnol. Bioeng. 113 1814–1824CrossRefGoogle Scholar
  13. Goldring MB, Tsuchimochi K and Ijiri K 2006 The control of chondrogenesis. J. Cell. Biochem. 97 33–44Google Scholar
  14. Govoni M, Berardi AC, Muscari C, et al. 2017 An engineered multiphase three-dimensional microenvironment to ensure the controlled delivery of cyclic strain and human growth differentiation factor 5 for the tenogenic commitment of human bone marrow mesenchymal stem cells. Tissue Eng. Part A 23 811–812CrossRefGoogle Scholar
  15. Grayson WL, Chao PH G, Marolt D, Kaplan DL and Vunjak-Novakovic G 2008 Engineering custom-designed osteochondral tissue grafts. Trends Biotechnol. 26 181–189Google Scholar
  16. Grayson WL, Marolt D, Bhumiratana S, Fröhlich M, Guo XE and Vunjak-Novakovic G 2011 Optimizing the medium perfusion rate in bone tissue engineering bioreactors. Biotechnol. Bioeng. 108 1159–1170Google Scholar
  17. Gullberg B, Johnell O and Kanis JA 1997 World-wide projections for hip fracture. Osteoporos. Int., 44 407–413CrossRefGoogle Scholar
  18. Hao J, Zhang Y, Jing D, et al. 2015 Mechanobiology of mesenchymal stem cells: perspective into mechanical induction of MSC fate. Acta Biomater. 20 1–9CrossRefGoogle Scholar
  19. Hoffmann W2015 Novel perfused compression bioreactor system as an in vitro model to investigate fracture healing. Front. Bioeng. Biotechnol. 3 1–6Google Scholar
  20. Huang C, Charles Y, Hagar KL, Frost LE, Sun Y and Cheung HS 2004 Effects of cyclic compressive loading on chondrogenesis of rabbit bone‐marrow derived mesenchymal stem cell. Stem Cells 22 313–323CrossRefGoogle Scholar
  21. Huh D, Hamilton GA and Ingber DE 2011 From 3D cell culture to organs-on-chips. Trends Cell Biol. 21 745–754CrossRefGoogle Scholar
  22. Izal I, Aranda P, Sanz-Ramos P, et al. 2013 Culture of human bone marrow-derived mesenchymal stem cells on of poly(l-lactic acid) scaffolds: potential application for the tissue engineering of cartilage. Knee Surgery Sports Traumatol. Arthrosc 21 1737–1750CrossRefGoogle Scholar
  23. Johnson RW, Schipani E and Giaccia AJ 2015 HIF targets in bone remodeling and metastatic disease. Pharmacol. Ther. 150 169–177CrossRefGoogle Scholar
  24. Kasper FK, Liao J, Kretlow JD, Sikavitsas VI and Mikos AG 2008 Flow perfusion culture of mesenchymal stem cells for bone tissue engineering; in StemBook (Cambridge, MA: Harvard Stem Cell Institute) pp 1–18Google Scholar
  25. Kleinhans C, Schmid FF, Schmid FV and Kluger PJ 2015 Comparison of osteoclastogenesis and resorption activity of human osteoclasts on tissue culture polystyrene and on natural extracellular bone matrix in 2D and 3D. J. Biotechnol. 205 101–110CrossRefGoogle Scholar
  26. Kock LM, Malda J, Dhert WJ A, Ito K and Gawlitta D2014 Flow-perfusion interferes with chondrogenic and hypertrophic matrix production by mesenchymal stem cells. J. Biomech. 47 2122–2129CrossRefGoogle Scholar
  27. Kumar G, Tison CK, Chatterjee K, et al. 2011 The determination of stem cell fate by 3D scaffold structures through the control of cell shape. Biomaterials 32 9188–9196CrossRefGoogle Scholar
  28. Le Nail LR, Stanovici J, Fournier J, Splingard M, Domenech J and Rosset P2014 Percutaneous grafting with bone marrow autologous concentrate for open tibia fractures: Analysis of forty three cases and literature review. Int. Orthopaedics 38 1845–1853Google Scholar
  29. Lee P, Mcaree M, Chang W and Yu X 2015 Mechanical forces in musculoskeletal tissue engineering. Regen. Eng. Musculoskeletal Tissues Interfaces CrossRefGoogle Scholar
  30. Liu C, Abedian R, Meister R, et al. 2012a Influence of perfusion and compression on the proliferation and differentiation of bone mesenchymal stromal cells seeded on polyurethane scaffolds. Biomaterials 33 1052–1064CrossRefGoogle Scholar
  31. Liu L, Yu B, Chen J, et al. 2012b Different effects of intermittent and continuous fluid shear stresses on osteogenic differentiation of human mesenchymal stem cells. Biomech. and Model. Mechanobiol. 11 391–401CrossRefGoogle Scholar
  32. Lohberger B, Kaltenegger H, Stuendl N, Payer M, Rinner B and Leithner A 2014 Effect of cyclic mechanical stimulation on the expression of osteogenesis genes in human intraoral mesenchymal stromal and progenitor cells. BioMed Res. Int. CrossRefGoogle Scholar
  33. Marijanovic I, Antunovic M, Matic I, Panek M and Ivkovic A 2016 Bioreactor-based bone tissue engineering. InTech Google Scholar
  34. Mekala NK, Baadhe RR and Parcha SR 2013 Study on osteoblast like behavior of umbilical cord blood cells on various combinations of PLGA scaffolds prepared by salt fusion. Curr. Stem Cell Res. Ther. 8 253–259CrossRefGoogle Scholar
  35. Mekala NK, Baadhe RR, Parcha SR and Prameela DY 2012 Osteoblast differentiation of umbilical cord blood-derived mesenchymal stem cells and enhanced cell adhesion by fibronectin. Tissue Eng. Regen. Med. 9 259–264CrossRefGoogle Scholar
  36. Mekala NK, Baadhe RR and Potumarthi R2014 Mass transfer aspects of 3D cell cultures in tissue engineering. Asia‐Pac. J. Chem. Eng. 9 318–329Google Scholar
  37. Mitra D, Whitehead J, Yasui OW and Leach JK 2017 Bioreactor culture duration of engineered constructs influences bone formation by mesenchymal stem cells. Biomaterials 146 29–39CrossRefGoogle Scholar
  38. Nishi M, Matsumoto R, Dong J and Uemura T2013 Engineered bone tissue associated with vascularization utilizing a rotating wall vessel bioreactor. J. Biomed. Mater. Res. Part A 101 421–427Google Scholar
  39. Ongaro A, Pellati A, Bagheri L, Fortini C, Setti S and De Mattei M2014 Pulsed electromagnetic fields stimulate osteogenic differentiation in human bone marrow and adipose tissue derived mesenchymal stem cells. Bioelectromagnetics 35 426–436Google Scholar
  40. Patterson JT, Gilliland T, Maxfield MW, et al. 2012 Tissue-engineered vascular grafts for use in the treatment of congenital heart disease: from the bench to the clinic and back again. Regen. Med. 7 409–419CrossRefGoogle Scholar
  41. Rauh J, Milan F, Günther K-P and Stiehler M 2011 Bioreactor systems for bone tissue engineering. Tissue Eng. Part B 17 263–280CrossRefGoogle Scholar
  42. Salzmann GM and Stoddart MJ 2014 Bioreactor tissue engineering for cartilage repair; in Emans P and Peterson L (eds) Developing Insights in Cartilage Repair (Springer, London) pp 79–97Google Scholar
  43. Sikavitsas VI, Bancroft GN, Lemoine JJ, Liebschner MA K, Dauner M and Mikos AG 2005 Flow perfusion enhances the calcified matrix deposition of marrow stromal cells in biodegradable nonwoven fiber mesh scaffolds. Ann. Biomed. Eng. 33 63–70CrossRefGoogle Scholar
  44. Sikavitsas VI, Bancroft GN and Mikos AG 2002 Formation of three-dimensional cell/polymer constructs for bone tissue engineering in a spinner flask and a rotating wall vessel bioreactor. J. Biomed. Mater. Res. 62 136–148CrossRefGoogle Scholar
  45. Stiehler M, Bünger C, Baatrup A, Lind M, Kassem M and Mygind T2009 Effect of dynamic 3-D culture on proliferation, distribution and osteogenic differentiation of human mesenchymal stem cells. J. Biomed. Mater. Res. Part A 89 96–107Google Scholar
  46. Tocchio A, Tamplenizza M, Martello F, et al. 2015 Biomaterials Versatile fabrication of vascularizable scaffolds for large tissue engineering in bioreactor. Biomaterials 45 124–131CrossRefGoogle Scholar
  47. Tutak W, Jyotsnendu G, Bajcsy P and Simon CG 2017 Nanofiber scaffolds influence organelle structure and function in bone marrow stromal cells. J. Biomed. Mater. Res. B Appl. Biomater. 105 989–1001CrossRefGoogle Scholar
  48. van den Dolder J, Bancroft GN, Sikavitsas VI, Spauwen PH M, Jansen JA and Mikos AG 2003 Flow perfusion culture of marrow stromal osteoblasts in titanium fiber mesh. J. Biomed. Mater. Res. Part A 64 235–241CrossRefGoogle Scholar
  49. Wang Y-K and Chen CS 2013 Cell adhesion and mechanical stimulation in the regulation of mesenchymal stem cell differentiation. J. Cell. Mol. Med. 17 823–832CrossRefGoogle Scholar
  50. Yang J, Cao C, Wang W, et al. 2010 Proliferation and osteogenesis of immortalized bone marrow-derived mesenchymal stem cells in porous polylactic glycolic acid scaffolds under perfusion culture. J. Biomed. Mater. Res. Part A 92 817–829Google Scholar
  51. Yeatts AB, Choquette DT and Fisher JP 2013 Bioreactors to influence stem cell fate: Augmentation of mesenchymal stem cell signaling pathways via dynamic culture systems. Biochim.t Biophys. Acta 1830 2470–2480CrossRefGoogle Scholar
  52. Yu H-S, Won J-E, Jin G-Z and Kim H-W 2012 Construction of mesenchymal stem cell–containing collagen gel with a macrochanneled polycaprolactone scaffold and the flow perfusion culturing for bone tissue engineering. BioResearch Open Access 1 124–136CrossRefGoogle Scholar
  53. Zermatten E, Vetsch JR, Ruffoni D, Hofmann S, Müller R and Steinfeld A2014 Micro-computed tomography based computational fluid dynamics for the determination of shear stresses in scaffolds within a perfusion bioreactor. Ann. Biomed. Eng. 42 1085–1094Google Scholar
  54. Zhang H, Mao X, Du Z, et al. 2016 Three dimensional printed macroporous polylactic acid/hydroxyapatite composite scaffolds for promoting bone formation in a critical-size rat calvarial defect model. Sci. Technol. Adv. Mater. 17 136–148CrossRefGoogle Scholar
  55. Zhang ZY, Teoh SH, Teo EY, et al. 2010 A comparison of bioreactors for culture of fetal mesenchymal stem cells for bone tissue engineering. Biomaterials 31 8684–8695CrossRefGoogle Scholar

Copyright information

© Indian Academy of Sciences 2019

Authors and Affiliations

  • Bhaskar Birru
    • 1
    Email author
  • Naveen Kumar Mekala
    • 2
  • Sreenivasa Rao Parcha
    • 1
  1. 1.Department of BiotechnologyNational Institute of TechnologyWarangalIndia
  2. 2.College of MedicineCentral Michigan UniversityMt PleasantUSA

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