Determination of Total Protein and Calcium in Gingival Mesenchymal Stem Cell-Conditioned Medium

Banun Kusumawardani, Intan Julita Purnamasari, Dea Ajeng Pravita Suendi

Abstract


Gingival mesenchymal stem cell-conditioned medium (GMSC-CM) is a conditioned medium obtained from cultured gingival mesenchymal stem cells. GMSCs are easily isolated from gingival tissue, whereas gingival tissue is easily obtained by minimally invasive techniques. On the other hand, CM contains proteins, cytokines, chemokines and growth factors that play an important role in osteogenic differentiation.This study aims to determine the total protein and calcium levels in GMSC-CM.GMSCs were grown in culture media with 10% of FBS. CM of GMSC was obtained from the media collection process with a 0.22 µm filter and concentrated with a centrifugal filter to obtain a concentrated GMSC-CM. The concentration of total protein was performed by bicinchoninic acid protein assay on GMSC-CM and concentrated GMSC-CM. Calcium level was performed by atomic absorption spectroscopy method. The results arethat GMSC-CM had a total protein concentration of 2502±0.06 µg/ml, and the concentratedGMSC-CM was 1.912±0.08 μg/ml.The calcium level of GMSC-CM was 0.009% and concentratedGMSC-CM was 0.009%. It can be concluded thatGMSC-CM had a high concentration of total protein and calcium. These parameters were used to develop bioprocesses to enhance the production of GMSC-CM, which will support the implementation of cell-free therapy for tissue regeneration.

Keywords


Calcium; Cell-free Therapy; Gingival Mesenchymal Stem Cell-conditioned Medium; Total Protein

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References


Kusumawardani B, Robin DMC, Puspitasari E, Savitri IJ, Suendi DAP. Cultivation and expansion of mesenchymal stem cells from human gingival tissue and periodontal ligament in different culture media. Dental Journal. 2021; 54(1): 39-45.

Vizoso FJ, Eiro N, Cid S, Schneider J, Perez-Fernandez R. Mesenchymal stem cell secretome: toward cell-free therapeutic strategies in regenerative medicine. International journal of molecular sciences. 2017; 18(9):1852.

Chaparro O, Linero I. Regenerative Medicine: A New Paradigm in Bone Regeneration. Advanced Techniques in Bone Regeneration. 2016; 253-74.

Pawitan JA. Prospect of stem cell conditioned medium in regenerative medicine. BioMed research international. 2014; 2014:1-14.

Katagiri W, Osugi M, Kawai T, Ueda M. Novel cell-free regeneration of bone using stem cell-derived growth factors. International Journal of Oral & Maxillofacial Implants. 2013; 28(4):1009-16.

Presen DM, Traweger A, Gimona M, Redl H. Mesenchymal stromal cell-based bone regeneration therapies: From cell transplantation and tissue engineering to therapeutic secretomes and extracellular vesicles. Frontiers in Bioengineering and Biotechnology. 2019; 7(352):1-20.

Kichenbrand C., Velot E., Menu P., Moby V. Dental Pulp Stem Cell-Derived Conditioned Medium: An Attractive Alternative for Regenerative Therapy. Tissue Eng. Part B Rev. 2018; 25(1):78-88.

Billing AM, Ben Hamidane H, Dib SS, Cotton RJ, Bhagwat AM, Kumar P, Hayat S, Yousri NA, Goswami N, Suhre K, Rafii A, Graumann J. Comprehensive transcriptomic and proteomic characterization of human mesenchymal stem cells reveals source specific cellular markers. Scientific Reports. 2016; 6(1):1-14.

Teixeira, F., Panchalingam, K., Assunção-Silva, R. et al. Modulation of the Mesenchymal Stem Cell Secretome Using Computer-Controlled Bioreactors: Impact on Neuronal Cell Proliferation, Survival and Differentiation. Scientific Reports. 2016; 6(1):1-14.

Pare B, Deschenes LT, Pouliot R, Dupre N, Gros-Louis F. An Optimized Approach to Recover Secreted Proteins from Fibroblast Conditioned-Media for Secretomic Analysis. Frontiers in Cellular Neuroscience. 2016; 10(70): 1-9.

Jeong HM, Jin Y-H, Choi YH, Yum J, Choi JK, Yeo C-Y, Lee K-Y. PKC signaling inhibits osteogenic differentiation through the regulation of Msx2 function. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. 2012; 1823(8):1225-32.

S. Taverna, M. Pucci, and R. Alessandro. Extracellular vesicles: small bricks for tissue repair/regeneration. Annals of Translational Medicine. 2017; 5(4):83.

Cut Yulian Fitriani, Awaluddin Wibawa. Titanium Biocompatibility on Dental Implant. Insisiva Dental Journal. 2019; 8(2): 53-58.

Y. Tachida, H. Sakurai, and J. Okutsu. Proteomic comparison of the secreted factors of mesenchymal stem cells from bone marrow, adipose tissue and dental pulp. Journal of Proteomics & Bioinformatics. 2015; 8(12):266–73.

P. R. Amable, M. V. T. Teixeira, R. B. V. Carias, J. M. Granjeiro, and R. Borojevic. Protein synthesis and secretion in human mesenchymal cells derived from bone marrow, adipose tissue and Wharton’s jelly. Stem Cell Research & Therapy. 2014; 5(2):1-13.

Lee MN, Hwang HS, Oh SH, Roshanzadeh A, Kim J-W, Song JH, Kim E-S, Koh J-T. Elevated extracellular calcium ions promote proliferation and migration of mesenchymal stem cells via increasing osteopontin expression. Experimental & Molecular Medicine. 2018; 50(11):1-16.




DOI: https://doi.org/10.18196/di.v10i2.13096

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