• Logo
  • SBMUJournals

Comparison of bone cell viability and proliferation in 3D scaffold to Monolayer cell culture

Faezeh Azizi, Sahar Omidpanah, Afshin Moradi, Mohammad Ali Hossini, Fereshte Aliakbari, Samira Shariatpanahi




Introduction: Today, due to high rates of accidents and fractures leading to bone defects and due to the limited possibility of bone graft bonding, using the patient’s cell culture on appropriate scaffolds and transferring it to the defect area is suggested as one of the treatment plans.

Materials and methods: Bone samples of 8 male subjects that were under craniotomy surgery in the hospital were collected. First, the samples were cut into smaller pieces and then, transferred to incubator culture dishes. Two weeks later, the osteoblast activity on the bone matrix began and on average, the cells covered the dishes within two weeks. The first generation of the cells was removed by Trypsin_EDTA method from the opaltes, then were divided into two parts, one was added to alginate gel and the other to monolayer culture. In order to prove the osteoblast activity on the bone matrix and investigate these activities, Van Kossa staining method was used, and also to investigate the cell viability, MTT method was employed.  

Results: There was a significant difference in the number of the cells created in alginate gel and those created in monolayer after two weeks (P <0.001). Moreover, the difference between mean cell counts in alginate gel and monolayer was statistically significant (P < 0.001). The results of the MTT test in second week showed that the number of alive cells is significantly higher in alginate gel (P <0.001). Finally, the result of the Van Kossa method proved extracellular matrix in both experimental groups.

Conclusion: Results showed that alginate gel better can support duplication and survival of osteoblasts compared to monolayer culture. This may be attributed to the biological properties of this gel; alginate gel porosity provides conditions under which cellular and metabolic activities are accelerated.



Williams C, Kadri OE, Voronov RS, Sikavitsas VI. Time-Dependent Shear Stress Distributions during Extended Flow Perfusion Culture of Bone Tissue Engineered Constructs. Fluids. 2018;3(2):25.

Hinze M, Sauerbier S, Wiedmann-Al-Ahmad M, Hübner U, Schmelzeisen R, Gutwald R. Bone Engineering: Allogenic and Alloplastic Bone Transplants vitalized by Osteoblast-like Cells. Issues. 2018;2017:2016.

Frohlich M, Grayson WL, Wan LQ, Marolt D, Drobnic M, Vunjak-Novakovic G. Tissue engineered bone grafts: biological requirements, tissue culture and clinical relevance. Current stem cell research & therapy. 2008;3(4):254-64.

Nguyen MK, Jeon O, Dang PN, Huynh CT, Varghai D, Riazi H, et al. RNA interfering molecule delivery from in situ forming biodegradable hydrogels for enhancement of bone formation in rat calvarial bone defects. Acta biomaterialia. 2018.

Silva GAB, Bertassoli BM, Sousa CA, Albergaria JD, de Paula RS, Jorge EC. Effects of strontium ranelate treatment on osteoblasts cultivated onto scaffolds of trabeculae bovine bone. Journal of bone and mineral metabolism. 2018;36(1):73-86.

Bose S, Roy M, Bandyopadhyay A. Recent advances in bone tissue engineering scaffolds. Trends in biotechnology. 2012;30(10):546-54.

Ishaug SL, Crane GM, Miller MJ, Yasko AW, Yaszemski MJ, Mikos AG. Bone formation by three‐dimensional stromal osteoblast culture in biodegradable polymer scaffolds. Journal of biomedical materials research. 1997;36(1):17-28.

Schantz J-T, Hutmacher DW, Lam CXF, Brinkmann M, Wong KM, Lim TC, et al. Repair of calvarial defects with customised tissue-engineered bone grafts II. Evaluation of cellular efficiency and efficacy in vivo. Tissue engineering. 2003;9(4, Supplement 1):127-39.

Sadeghi H, Bahramian H, Hashemibeni B, Esfandiary E, Aliakbari F. Comparison of Human Osteoblast Proliferation in Alginate and Hydroxyapatite-Tricalcium Phosphate Scafolds. Journal of Isfahan Medical School. 2010;28(111).

Wiesmann HP, Nazer N, Klatt C, Szuwart T, Meyer U. Bone tissue engineering by primary osteoblast-like cells in a monolayer system and 3-dimensional collagen gel. Journal of oral and maxillofacial surgery. 2003;61(12):1455-62.

Hashemibeni B, Sadeghi F, Bahrambeigi V, Sanaei M, Sharifi E, Valiani A, et al. Determination and comparison rate of expression markers of osteoblast derived of Adipose derived stem cells markers in monolayer and pellet culture models. International Journal of Engineering Research and Development. 2015;11(1).

Chen C-Y, Ke C-J, Yen K-C, Hsieh H-C, Sun J-S, Lin F-H. 3D porous calcium-alginate scaffolds cell culture system improved human osteoblast cell clusters for cell therapy. Theranostics. 2015;5(6):643.

Venkatesan J, Bhatnagar I, Manivasagan P, Kang K-H, Kim S-K. Alginate composites for bone tissue engineering: a review. International journal of biological macromolecules. 2015;72:269-81.

Lin HY, Chang TW, Peng TK. Three‐dimensional plotted alginate fibers embedded with diclofenac and bone cells coated with chitosan for bone regeneration during inflammation. Journal of Biomedical Materials Research Part A. 2018;106(6):1511-21.

Wang MO, Bracaglia L, Thompson JA, Fisher JP. Hydroxyapatite‐doped alginate beads as scaffolds for the osteoblastic differentiation of mesenchymal stem cells. Journal of Biomedical Materials Research Part A. 2016;104(9):2325-33.

Gutiérrez-Hernández JM, Escobar-García DM, Escalante A, Flores H, González FJ, Gatenholm P, et al. In vitro evaluation of osteoblastic cells on bacterial cellulose modified with multi-walled carbon nanotubes as scaffold for bone regeneration. Materials Science and Engineering: C. 2017;75:445-53.

Maes C, Kobayashi T, Selig MK, Torrekens S, Roth SI, Mackem S, et al. Osteoblast precursors, but not mature osteoblasts, move into developing and fractured bones along with invading blood vessels. Developmental cell. 2010;19(2):329-44.

Pape H-C, Lehmann U, Van Griensven M, Gänsslen A, Von Glinski S, Krettek C. Heterotopic ossifications in patients after severe blunt trauma with and without head trauma: incidence and patterns of distribution. Journal of orthopaedic trauma. 2001;15(4):229-37.

Gómez-Barrena E, Rosset P, Lozano D, Stanovici J, Ermthaller C, Gerbhard F. Bone fracture healing: cell therapy in delayed unions and nonunions. Bone. 2015;70:93-101.

Coelho M, Cabral AT, Fernandes M. Human bone cell cultures in biocompatibility testing. Part I: osteoblastic differentiation of serially passaged human bone marrow cells cultured in α-MEM and in DMEM. Biomaterials. 2000;21(11):1087-94.

Hamlet SM, Vaquette C, Shah A, Hutmacher DW, Ivanovski S. 3‐Dimensional functionalized polycaprolactone‐hyaluronic acid hydrogel constructs for bone tissue engineering. Journal of clinical periodontology. 2017;44(4):428-37.

Klotz BJ, Gawlitta D, Rosenberg AJ, Malda J, Melchels FP. Gelatin-methacryloyl hydrogels: towards biofabrication-based tissue repair. Trends in biotechnology. 2016;34(5):394-407.

Sarker B, Rompf J, Silva R, Lang N, Detsch R, Kaschta J, et al. Alginate-based hydrogels with improved adhesive properties for cell encapsulation. International journal of biological macromolecules. 2015;78:72-8.

Fan C, Wang D-A. Macroporous hydrogel scaffolds for three-dimensional cell culture and tissue engineering. Tissue Engineering Part B: Reviews. 2017;23(5):451-61.

Abbah S, Lu W, Chan D, Cheung K, Liu W, Zhao F, et al. Osteogenic behavior of alginate encapsulated bone marrow stromal cells: an in vitro study. Journal of Materials Science: Materials in Medicine. 2008;19(5):2113-9.

Liao C-C, Kao M-C. Cranioplasty for patients with severe depressed skull bone defect after cerebrospinal fluid shunting. Journal of clinical neuroscience. 2002;9(5):553-5.

Alsberg E, Anderson K, Albeiruti A, Franceschi R, Mooney D. Cell-interactive alginate hydrogels for bone tissue engineering. Journal of dental research. 2001;80(11):2025-9.


  • There are currently no refbacks.