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:: Volume 27, Issue 6 (Scientific Journal of Kurdistan University of Medical Sciences 2023) ::
SJKU 2023, 27(6): 25-36 Back to browse issues page
Optimizing a Calorimetric Method for Quantitative Assessment of Alkaline Phosphatase to Confirm the Osteogenic Differentiation of Mesenchymal Stem Cell
Samaneh Harimi1 , Fatemeh Zare2 , Farzaneh Fesahat2 , Mahdieh Mondanizadeh 3
1- Master of Medical Biotechnology, Student Research Committee, Arak University of Medical Sciences, Arak, Iran
2- Assistant Professor, Reproductive Immunology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
3- Associate Professor, Department of Biotechnology and Molecular Medicine, School of Medicine, Arak University of Medical Sciences, Arak, Iran , m_mondanizadeh@yahoo.com
Abstract:   (1236 Views)
Background and Aim: Evaluation of the alkaline phosphatase expression is known as one of the diagnostic markers to confirm the stem cells differentiation into osteoblasts. Several methods, including immunocytochemistry, are used to measure alkaline phosphatase in different studies. Due to limitations of using this method, we decided to optimize the calorimetric method as an alternative, low cost and efficient method for quantitative assessment of alkaline phosphatase.
Materials and Methods: Mesenchymal stem cells derived from human adipose tissue were used. Cells were differentiated into osteoblasts using an induction medium. Alkaline phosphatase levels were assessed on the days 0 and 21 of differentiation using both immunocytochemistry and calorimetric methods.
Results: Immunocytochemistry findings demonstrated a significant increase (approximately 5-fold) in alkaline phosphatase expression level on the 21st day of differentiation compared to that on the day 0 (p <0.0001). Also, statistical analysis of the results of the calorimetric test showed a significant increase (approximately 9-fold) in alkaline phosphatase expression level on the 21st day of differentiation compared to  that on the day 0 (p<0.0001).
Conclusion: Comparison of the data between calorimetric and immunocytochemistry showed similar results. These findings suggested that colorimetric assay can be used as an alternative, quantitative, fast, and low cost method for determining the levels of alkaline phosphatase in osteoblasts differentiated from mesenchymal stem cells.
 
Keywords: Alkaline phosphatase, Differentiation, Stem cells, Osteoblast, Calorimetric method
Full-Text [PDF 1502 kb]   (519 Downloads)    
Type of Study: Original Research | Subject: Biotechnology
Received: 2022/01/16 | Accepted: 2022/06/21 | Published: 2023/03/3
References
1. Oryan A, Monazzah S, Bigham-Sadegh A. Bone injury and fracture healing biology. Biomed Environ. Sci. 2015;28(1):57-71.
2. Gong Y, Yang J, Li X, Zhou C, Chen Y, Wang Z, et al. A
3. dissection of human primary osteoblasts in vivo at single-cell resolution. Aging (Albany NY). 2021;13(16):20629. [DOI:10.18632/aging.203452] [PMID] []
4. Asserson DB, Orbay H, Sahar DE. Review of the pathways involved in the osteogenic differentiation of adipose-derived stem cells. J Craniofac Surg. 2019;30(3):703-8. [DOI:10.1097/SCS.0000000000005447] [PMID]
5. Sharma U, Pal D, Prasad R. Alkaline phosphatase: an overview. Indian J Clin Biochem. 2014;29(3):269-78. [DOI:10.1007/s12291-013-0408-y] [PMID] []
6. Jafary F, Hanachi P, Gorjipour K. Osteoblast differentiation on collagen scaffold with immobilized alkaline phosphatase. IJOTM. 2017;8(4):195.
7. Lowe D, Sanvictores T, John S. Alkaline phosphatase. StatPearls Publishing, Treasure Island (FL);2017:95-97.
8. Sebastián-Serrano Á, de Diego-García L, Martínez-Frailes C, Ávila J, Zimmermann H, Millán JL, et al. Tissue-nonspecific alkaline phosphatase regulates purinergic transmission in the central nervous system during development and disease. Comput Struct Biotechnol J. 2015;13:95-100. [DOI:10.1016/j.csbj.2014.12.004] [PMID] []
9. Im K, Mareninov S, Diaz MFP, Yong WH. An introduction to performing immunofluorescence staining. Yong, W. (eds)Biobanking. Methods in Molecular Biology, vol 1897. Humana Press, New York, NY. 2019:299-311. [DOI:10.1007/978-1-4939-8935-5_26] [PMID] []
10. Burry RW. Controls for immunocytochemistry: an update. J Histochem Cytochem.2011;59(1):6-12. [DOI:10.1369/jhc.2010.956920] [PMID] []
11. Boghori M, Aghamaali M, Sariri R, Mohamadpour F, Ghafouri H. Salivary enzymes and flow rate: Markers of peptic ulcer. J Oral Biol Craniofac Res. 2014;4(1):24-9. [DOI:10.1016/j.jobcr.2013.12.001] [PMID] []
12. Folgiero V, Migliano E, Tedesco M, Iacovelli S, Bon G, Torre ML, et al. Purification and characterization of adipose-derived stem cells from patients with lipoaspirate transplant. Cell Med. 2010;1(1):3-14. [DOI:10.3727/215517910X519265]
13. Arinzeh TL. Mesenchymal stem cells for bone repair: preclinical studies and potential orthopedic applications. Foot Ankle Clin. 2005;10(4):651-65. [DOI:10.1016/j.fcl.2005.06.004] [PMID]
14. Arpornmaeklong P, Brown SE, Wang Z, Krebsbach PH. Phenotypic characterization, osteoblastic differentiation, and bone regeneration capacity of human embryonic stem cell-derived mesenchymal stem cells. Stem cells dev. 2009;18(7):955-68. [DOI:10.1089/scd.2008.0310] [PMID] []
15. Subramani K, Pandruvada S, Puleo D, Hartsfield J, Huja S. In vitro evaluation of osteoblast responses to carbon nanotube-coated titanium surfaces. Prog Orthod. 2016;17(1):1-9. [DOI:10.1186/s40510-016-0136-y] [PMID] []
16. Li W, Zhang S, Liu J, Liu Y, Liang Q. Vitamin K2 stimulates MC3T3‑E1 osteoblast differentiation and mineralization through autophagy induction. Mol Med Rep. 2019;19(5):3676-84. [DOI:10.3892/mmr.2019.10040] [PMID] []
17. Oryan A, Alidadi S, Moshiri A. Current concerns regarding healing of bone defects. Hard tissue. 2013;2(2):1-12. [DOI:10.13172/2050-2303-2-2-374]
18. Rilianawati R, Bratakencana J, Harlim A. Differentiation Potential of Adipose-Derived Mesenchymal Stem Cells to Osteoblast Cell in Early, Middle and Late Passages. Stem Cell Res Ther. 2018;8(5):2-7.
19. Grottkau BE, Lin Y. Osteogenesis of adipose-derived stem cells. Bone res. 2013;1(1):133-45. [DOI:10.4248/BR201302003] [PMID] []
20. Hanna H, Mir LM, Andre FM. In vitro osteoblastic differentiation of mesenchymal stem cells generates cell layers with distinct properties. Stem Cell Res Ther. 2018;9(1):1-11. [DOI:10.1186/s13287-018-0942-x] [PMID] []
21. OakáKeem J, HyunáChung B. Smart nanoprobes for the detection of alkaline phosphatase activity during osteoblast differentiation. ChemComm. 2015;51(15):3270-2. [DOI:10.1039/C4CC09620G] [PMID]
22. Osathanon T, Giachelli CM, Somerman MJ. Immobilization of alkaline phosphatase on microporous nanofibrous fibrin scaffolds for bone tissue engineering. Biomaterials. 2009;30(27):4513-21. [DOI:10.1016/j.biomaterials.2009.05.022] [PMID] []
23. Birmingham E, Niebur G, McHugh PE. Osteogenic differentiation of mesenchymal stem cells is regulated by osteocyte and osteoblast cells in a simplified bone niche. Eur Cells Mater. 2012;23:13-27. [DOI:10.22203/eCM.v023a02] [PMID]
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Harimi S, Zare F, Fesahat F, Mondanizadeh M. Optimizing a Calorimetric Method for Quantitative Assessment of Alkaline Phosphatase to Confirm the Osteogenic Differentiation of Mesenchymal Stem Cell. SJKU 2023; 27 (6) :25-36
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Volume 27, Issue 6 (Scientific Journal of Kurdistan University of Medical Sciences 2023) Back to browse issues page
مجله علمی دانشگاه علوم پزشکی کردستان Scientific Journal of Kurdistan University of Medical Sciences
مجله علمی دانشگاه علوم پزشکی کردستان Scientific Journal of Kurdistan University of Medical Sciences
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