Secretion of osteocalcin in chitosan–hydroxyapatite scaffold with seeding of cryopreserved human adipose derived mesenchymal stem cells

Background: Tissue engineering has developed in recent years, including the utilization of bone graft to regenerate bone. The incorporation of chitosan and hydroxyapatite as scaffolds improves osteoconductive ability. Human adipose derived mesenchymal stem cells (hADMSCs) potentially differentiate into osteoblasts and can accelerate the osteogenesis process. 
Purpose: To determine osteocalcin secretion on chitosan-hydroxyapatite (CS-HA) scaffold with seeding of cryopreserved hADMSCs in vitro
Method: Thawing was conducted on frozen (hADMSCs) and characterized using immunocytochemistry. HADMSCs were seeded on CS-HA scaffolds. Examination of osteocalcin secretion levels was carried out by collecting cell culture supernatants of hADMSCs on 7th, 14th and 21st day of the observation group by using osteocalcin ELISA kit. 
Result: Scanning observation (SEM) proved that the chitosan-hydroxyapatite scaffold has a suitable pore structure, because hADMSCs can attach and differentiate when attached to the porosity of the scaffold. There were significant differences in osteocalcin secretion on days 7, 14 and 21 according to the ELISA results. 
Conclusion: The presence of bone regeneration process in chitosan-hydroxyapatite scaffold. This was characterized by significant osteocalcin secretion on 7th, 14th and 21st day. So it can be concluded that the scaffold is compatible.