| Objective: To explore the effects of treatment of platelet-rich plasma(PRP)united with Shaoyang Shenggu prescription on knee cartilage defect in rats by in vivo experiment.Methods: 1.Experimental grouping: 20 clean SD rats were divided into four groups according to random number table,A,B,C and D,with 5 rats in each group.A stands for control group,B for Shaoyang Shenggu prescription group,C for PRP group and D for PRP combined with Shaoyang Shenggu prescription group.2.Blood collection and PRP preparation: 3ml venous blood was taken from the posterior orbital vein of rats in Group C and Group D,PRP was prepared by secondary centrifugation.3.Model preparation and postoperative intervention: The animal model of full-thickness articular cartilage defect was established by physical injury in the middle of femoral trochlea of right knee of rats.Group A: cartilage defect area and postoperative non-treatment;Group B: after operation,Shaoyang Shenggu Decoction was given to the stomach.Group C: PRP was injected into articular cavity after operation;Group D: PRP was injected into articular cavity after operation and Shaoyang Shenggu Decoction was given to stomach after operation.4.Observation index: All rats were killed at the 3rd month after operation and cartilage tissue was roughly observed.Pathological tissue was detected by HE and toluidine blue staining,Mankin score was used to evaluate the repair of articular cartilage after injury.Ⅰmmunohistochemical method to observe the expression of Col Ⅰ and Col Ⅱ to evaluate the repair of articular cartilage.The positive expression of type Ⅱ collagen fiber was analyzed by average optical density to evaluate the repair effect of damaged cartilage.Results: 1.Gross specimen observation showed that the boundary between the defect area and the surrounding normal area in Group A was obvious,with large defect surface,uneven edge and partial calcification area,no obvious cartilage tissue repair was found;In group B and C,the boundary between the damaged area and the surrounding normal area is obvious,the damaged cartilage can be filled with smooth milky white tissue,but the damaged area is not filled;In group D,the boundary between the defect area and normal cartilage is not obvious.The damaged cartilage was filled with nearly transparent tissue with flat surface,and the damaged area was nearly filled.2.Histopathological staining showed that the chondrocytes in the repaired tissue of Group A were disordered and few in number,there was no obvious secretion of normal articular cartilage matrix,but a small amount of type Ⅰ collagen and type Ⅱ collagen were expressed.In group B,chondrocytes are scattered and the number of cells is small,with a small amount of purple-blue granules,a small amount of cartilage matrix secretion,rough cartilage surface and more type Ⅱ collagen expression.In group C,the cells in the repaired tissue were scattered and few in number,with a small amount of purple-blue particles and a small amount of matrix secreted,the cartilage surface was rough and more type Ⅱ collagen was expressed In group D,the number of cells in the repaired tissue increased significantly,and the cells appeared in clusters,showing cartilage lacunae,a large number of purple-blue granules,secretion of nearly normal cartilage matrix,and expression of type Ⅱ collagen increased significantly,showing brown-yellow or brown-brown granules.The new tissue in the defect area was well connected with the nearby cartilage tissue,the repaired cartilage surface was smooth and flat.3.The scores of Mankin in A,B,C and D groups were 12.60±0.55,10.40±0.55,10.60±0.55 and 8.40±0.55,respectively.The scores of Mankin in PRP combined with Shaoyang Shenggu Prescription group were significantly lower than those in control group and control group Mankin scores of Shaoyang Shenggu Prescription group and PRP group were obviously below those of control group(P < 0.001).Less than obvious difference between Shaoyang Shenggu Prescription group and PRP group(P > 0.05).4.The average optical density of type Ⅱ collagen expression showed that the average optical density of samples in A,B,C and D groups were 0.2469 ±0.0048,0.2566±0.0058,0.2597±0.0035 and 0.2761±0.0055,respectively.The mean optical density of PRP united with Shaoyang Shenggu Prescription group was more than that of other group.The mean optical density of Shaoyang Shenggu Prescription group and PRP group was more than that of control group,the difference was obvious significant(P < 0.05).Less than significant difference between Shaoyang Shenggu Prescription group and PRP group(P >0.05).Conclusion: The combination of autologous platelet-rich plasma and Shaoyang Shenggu Prescription can improve the curative effect of treating articular cartilage defects in rats and provide a new idea for regeneration and repair of cartilage injury. |
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