An in vitro study assessing the effects of methyl prednisolone and hyaluronic acid on human chondrocyte cell metabolism.

Presenting Author - George Jacob
Background: Conservative modalities such as intra articular corticosteroid and hyaluronic acid (HA) have been employed to ameliorate the symptoms of osteoarthrosis and delay total knee arthroplasty. They have both been used for decades with ambiguous results in literature. We studied the effects of the concentration of methyl prednisolone, low molecular weight (LMW) and high molecular weight (HMW) HA on human chondrocyte cell count, viability and CD44+ expression. Materials & Methods All procedures were approved under the institutional ethical committee and with patient informed consent. Three cartilage bone plugs were harvested during total knee arthroplasty from patients suffering from osteoarthritis scheduled for surgery under sterile conditions. Chondrocytes were isolated and cultured to passage one, three samples from passage one were cultured to passage two. Both passage chondrocytes were seeded in T-25 flasks with concentrations of 1 uM/ml, 10 uM/ml & 100 uM/ml of methyl prednisolone and 0.1 mg/ml, 1mg/ml & 2 mg/ml HMW and LMW HA against control. Regular medium changes were done and cells harvested on day 14 for assessment. Cell viability: after necessary slide preparation using a hemocytometer cells were counted in 4 squares under 100X magnification. Stained cells (non viable) and non stained cells (viable) were counted and made into a percentage. Cell count: assessed using a coulter counter after slide preparation. CD44+ expression: measured using flowcytometry Statistics: Statistical analysis was done with ANOVA test and independent t test, a P value of 0.05 was taken to be significant. Results: Cell count in the both passages decreased with increasing methylprednisolone concentration. The decrease in cell count in passage 2 was noted to be significant (P=0.013). Cell viability reduced in both passages with increased concentration of methylprednisolone with the decrease in passage 2 being significant (P=0.011). CD44+ expression decreased with increasing methyl prednisolone concentration in both passages, but the reduction was not significant. As the concentration of both HMW and LMW HA increases, there is decrease in cell viability count and CD44+ expression in both passage 1 and 2 but the decrease is not statistically significant. However, the decrease in CD44+ count in HMW HA passage 1 is significant (p value=0.04). Conclusion: Increasing concentrations of methyl prednisolone and both hyaluronic acid variants cause a decrease in cell viability, count and CD44+ expression. Our study indicates the use of methylprednisolone and HA do not promote chondrocyte proliferation and aid in treatment of osteoarthrosis and therefore are not effective treatment options.