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

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.