1.Preventive effect of No. Ⅰ recipe for osteoarthritis on chondrocyte injury by hydrocortisone in rabbits
Chinese Journal of Tissue Engineering Research 2005;9(38):190-192
BACKGROUND:Kidney-nourishing and blood activating Chinese herbs in the No. Ⅰ recipe for ostooarthritis have the effects of treating osteoarthritis and preventing the side effect of glucocorticoid.OBJECTIVE: To build up rabbit chondrocyte culture system in vitro to investigate the effect of preventive effect of No. Ⅰ recipe for osteoarthritis on chondrocyte injury by hydrocortisone.DESIGN:A complete randomized design and controlled trial.SETTING: Department of Rheumatology and Immunology, Second Hospital of Fujian ProvinceMATERIALS:The experiment was conducted from November 2003 to March 2004 at the Molecular Laboratory of Department of Orthopedics &Grammatology, Fujian College of Traditional Chinese Medicine. No. Ⅰrecipe for osteoarthritis consisted of Herba Epimedii, Radix Morindae Officinalis, Rhizoma Ligustic Chuanxiong, Radix Notoginseng and Daphne geraldii Nitsche. Articular cartilage was digested with collagenase Ⅱ ,chondrocyte culture system in vitro was built up, then in the culture system the different concentrations of the mixture of No. Ⅰ recipe for osteoarthritis and hydrocortisone were added, according to the concentrations they were at random divided into two groups. One group was added 1.25 g/L hydrocortisone and/or 150 g/L No. Ⅰ recipe for osteoarthritis, the other group was added 2.50 g/L hydrocortisone and/or 300 g/L No. Ⅰ recipe for osteoarthritis. Each group was again subdivided as control, hydrocortisone, hydrocortisone + No. Ⅰ recipe for osteoarthritis and No. Ⅰ recipe for osteoarthritis groups, with 5 wells in each subgroup.teoarthritis group: In each well 0.1 mL mixture of 1.25 g/L hydrocortihydrocortisone group: In each well 0.1 mL mixture of phosphate buffer teoarthritis group: In each well No. 1 recipe for osteoarthritis 0.1mL was In each well 0.1 mL mixture of 2.50 g/L hydrocortisone and 300 g/L No.each well 0.1mL mixture of phosphate buffer and 2.50 g/L hydrocortisone after medicines were added, the morphological change of chondrocyte in all (A) value in each well was assayed with tetrazoline colorimetry as reflection of proliferation of the chondrocyte. MAIN OUTCOME MEASURES:Influence of No. Ⅰ recipe for osteoarthritis combined with hydrocortisone on chondrocyte proliferation in rabbits.croscope that the chondrocytes in the control group were mostly in multiangles or shuttle-like, and adhesive growth; those cultured for 24 hours with 1.25 and 2.50 g/L hydrocortisone were in pyknosis or flotation state.;those in the various concentrations of hydrocortisone +No. Ⅰ recipe for osteoarthritis, and No. Ⅰ recipe for osteoarthritis groups were mostly in multiangles or shuttle-like, and adhesive growth, without difference with those in 1.25 g/L hydrocortisone group than in the control group(0.007 6±0.0018,0.015 2±0.002 6,t=5.374 0,P<0.01);it was obviously higher in the 1.25 g/L hydrocortisone +150 g/L .No. Ⅰ recipe for osteoarthritis and 150 g/L No. Ⅰ recipe for osteoarthritis groups than in the 1.25 g/L hydrocortisone group (0.065 6±0.016 9, 0.086 6±0.019 0, t=7.630 9, 9.255 9,P<0.01); it was obviously lower in the 2.50g/L hydrocortisone group than in the control group 2(0.005 3±0.001 4,0.015 3±0.002 3 ,t=8.304 5,P<0.01) ;it was obviously higher in the 2.50 g/L hydrocortisone +300 g/L No. Ⅰ recipe for osteoarthritis and 300 g/L No. Ⅰ recipe for osteoarthritis groups than in the 2.50 g/L hydrocortisone group (0.085 8±0.015 0,0.092 6 ±0.016 8, t=11.948 3,11.579 4, P < 0.01).CONCLUSION :Hydrocortisone in higher concentration could cause serious injury or death of chondrocytes, however the chondrocyte grew well cultured in the mixture of hydrocortisone and traditional Chinese drug, No.Ⅰ recipe for osteoarthritis had the effect of preventing injury of chondrocyte from hydrocortisone.