Hyaluronan promote chondrogenesis of human mesenchymal stem cell in tricalcium phosphate-collagen scaffolds

Abstract

Objective To investigate the effect of hyaluronan (HA) on engineered cartilage formation.Methods Human mesenchymal stem cell (hBMSCs) were isolated by density gradient centrifugation and were seeded onto tricalcium phosphate-collagen (TCP-COL) scaffolds.hBMSCs were cultured on the scaffold for 2 weeks in two groups:(1) control group ; (2) HA group (control group culture medium with 100 mg/L HA).The effect of hyaluronan on chondrogenesis was assessed by hematoxylin-eosin (HE) staining,alcian blue staining,anti type Ⅱ collagen immunohistochemical staining,Real-time polymerase chain reaction (RT-PCR) and glycosaminoglycans (GAG) quantification assay.Results The data of hematoxylin-eosin (HE) staining,alcian blue staining,anti type Ⅱ collagen immunohistochemical staining,RT-PCR [Collagen Ⅱ (COL2) (P ＜ 0.01),SRY-related high mobility group-box gene9 (SOX9) (P ＜ 0.01),Aggrecan (P ＜ 0.05)] (8.954 ± 2.612/1.062 ± 0.459,3.512 ± 0.638/1.111 ± 0.595 and 4.588 ± 1.964/1.042 ± 0.335,respectively) and GAG quantification assay (21.741 ± 2.633/16.717 ± 0.595) (P ＜ 0.05) shows HA (100 g/L) could significantly induce the chondrogenic differentiation of hBMSCs on the TCP-COL scaffold.In addition,in the HA group,a significant up-regulation of COL1 (P＜0.05) (3.258 ±1.038/1.127 ±0.678) andCOL10 (P＜0.05) (5.961 ±2.598/1.080 ±0.458) gene were detected compared with the control group,indicating that HA may also promote robust maturation of tissue-engineered cartilage.Conclusion HA not only could significantly induce the chondrogenic differentiation of hBMSCs on the TCP-COL scaffolds,but also could promote maturation of the tissue-engineered cartilage. Key words: Hyaluronic acid; Bone marrow mesenchymal stem cells; Tissue-engineering cartilage