Secretion of osteocalcin in chitosan–hydroxyapatite scaffold with seeding of cryopreserved human umbilical cord mesenchymal stem cells

Michael Josef Kridanto Kamandjadja 1, *, Erike Dwi Safitri 2, Patricya Talitha Damayanti 2 and Shima Ayu Nourma Kholifah 3

1 Department of Prosthodontics, Faculty of Dental Medicine, Airlangga University, Surabaya, Indonesia.
2 Undergraduate, Faculty of Dental Medicine, Airlangga University, Surabaya, Indonesia.
3 Stem Cells Research Center and Development, Institute Tropical Disease, Airlangga University, Surabaya, Indonesia.
 
Research Article
World Journal of Advanced Research and Reviews, 2024, 21(01), 288–302
Article DOI: 10.30574/wjarr.2024.21.1.2625
 
Publication history: 
Received on 12 November 2023; revised on 21 December 2023; accepted on 23 December 2023
 
Abstract: 
In recent years, regenerative therapy using tissue engineering has rapidly developed. One of the applications is for alveolar bone tissue engineering in prosthodontics. Currently, the most widely developed biomaterial in bone tissue engineering is a scaffold. The incorporation of chitosan and hydroxyapatite (CS-HA) improves osteoconductive ability. Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) are capable of acting as osteoprogenitors and promoting bone formation. Purpose: To determine the influence of osteocalcin secretion in the CS[1]HA scaffold with seeding of cryopreserved hUCMSCs in vitro. Methods: The thawing process was carried out on frozen hUCMSCs, which were then examined for characterization using immunocytochemistry. The hUCMSCs were seeded on the CS-HA scaffold. Finally, examination of osteocalcin secretion levels was carried out by collecting cell culture supernatant on the 14th and 21st days of observation groups using the human osteocalcin ELISA kit. Results: Immunocytochemistry testing revealed that the hUCMSCs had the characteristics of the MSCs marker, namely positive expression on the CD90 marker and negative expression on the CD45 marker. Scanning electron microscope (SEM) observation revealed that the CS-HA scaffold had a structure with many pores and hUCMSCs could be attached and proliferation among the porosity of the scaffold. Based on the ELISA result, there was a significant difference in the average amount of osteocalcin secretion on days 14 and 21 which indicated osteogenic differentiation. Conclusion: Osteocalcin secretion in the CS-HA scaffold using the seeding of in vitro hUCMSCs demonstrated a bone regeneration process that showed a significant increase on day 21.
 
Keywords: 
Bone Tissue Engineering; Scaffold Chitosan-Hydroxyapatite; Osteocalcin; Human Umbilical Cord Mesenchymal Stem Cells
 
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