Histological Observation Of BFGF Influences On Intrinsic Healing Of Flexor Tendons Within Synoviail Sheath

Objective: We often see injury of tendon in clinical orthopae- dics especially flexor tendons injury. It has always been a difficult problom in healing and adhesion after operating. Doctors try to recover its continuous, keep recovering, try to solve the injured smooth, but it always takes most of the patients a lot of time for external fixation, and recover the function with secondary tendonlysis, but we cannot always get efficacy well. For many years, people do a lot of experimental research on anatomy, physiological, pathological and the process of the healing. They explored many good ways to promote the healing of tendons and prevention adhesion. They have made some achievement. However, we still cannot find a good way, and a cheaper way to develop in clinical.bFGF is a member of heparin binding growth factor family. It consists of 146 amino acid residues, and is a polypeptide. Studies have shown that exogenous bFGF not only can promote the healing of tendons, but also can facilitate the formation of tendon adhesion. It is a very important that choose the proper way so as to developing merits, avoiding weakness. It has been the main point to study at home and abroad. So far, we do the experiment to make the venous indwelling trocar to break end. We continus injecting a little bFGF early time, intervent the process of the healing of tendon. To investigate the effect of bFGF influences on intrinsic healing of flexor tendons within synoviail sheath from histological observation, and compared the processes of healing and adhesion. To provide theoretical foundation that promote the process of healing of tendon after injuring, make it fits for early stage dirigation.Methods: We used 78 healthy male leghorns, divided leg- horn into 3 groups randomly, each chicken was anaesthesiaed with Ketamine intramuscular injection, and then fixed on the operating board . The right foot was disinfected routily. Cut the skin of proximal phalanx and midplate of the third dactly as "┏┓" shape separates the tela subcutanea, cuts open the sheath tube between the A2-A4 pulley. After the model of the deep toe flexor tendon breaking cross had been made, proximal longitudinal near group A put the venous indwelling needle, opened at the break end, in order to put the medicine in it. Within flexor sheath on group B put the venous indwelling needle, in order to put medicine in flexor sheath, suture in situ on the original of group C, make it normal healing. With 6-0 non-injured suture line, using the method of Modified Kessler suture technique after the tendon had been severed. Then with 7-0 non-injured suture line to suture the sheath, use the plaster to fix chicken's foot as boxing gloves. After operation, the chicken is raised in the cage free-running, using the antibiotics for 3 days to prevent infection, injected 5.0ng/ml of bFGF after opration 24 hours, 2ul for group A once a time, 5ul for group B once a time, twelve hours once, go on for 8 times, then pull the indwelling needle out after 12 hours of the last one. 2,3,4,6 weeks after the operation, six toes of leghorns in every group were dissected layer-by-layer in local anesthetic. The conditions of tendon healing and adhesion were observed. Every section was used for HE staining, Masson's staining, and the conditions of tendon were observed under light microscope, the function of cells was observed under transmission electron microscope (TEM) and scanning electron microscope (SEM).The picture analysis of the sections was dealt with cell parameter analysis system. Put the data into computer to statistical analysis.Result: Gross appearance: Two weeks after operating, translucent colloidal of each group exists in the break end and the around. The tendon break end in group B was wider adhesion, a bit red, more looser, connective tissue filled in between the tendon sheath and tendon, it is crisp. The adhesion of A and C is a little lighter than group B, the substance of colloudal in group B is similar to group A, it is a little darker than group C in color, the toughness is stronger than group C; Three weeks after operating, the color of the shuttle-shape connection is darker and less crisp, the adhesion was more seriously, the scope of adhesion in group B is wider, and the blunt seperation is harder than before, the break end connected stronger and the tendon gliding movement passively is limited. In group A and C, only the shuttle-shape connection adhered, the blunt seperation was easier than group B; The color of break end colloidal in every group was more epinepheloser, the toughness is more stranger, but group C is weaker. Four weeks after operating, the pyknotic tendon adhesion were formed in group B, the adhesion band was the same as that before one week, and mast be divided by sharp separation. The break end was connected firmly by fibrous tissues, gray in color, pliable but strong, and elasticity was good. Adhension between A and C was very similar, but it is clear that the quality of healing of group A was better than group C. Six weeks after operating, the degree of adhension of group A and C were similar, more lighter than group B, the quality of healing of group A and group B were similar, and it was better than group C.Hisological observation: The break end between group A and group B are similar, the adhension group A and group C was nearly the same. Two weeks after operating, the fibrolasts in tendon group A proliferate very activity so we can find new collagen fibers and hyperplasia of capillaries, obviously more than peri-tendon, he fibrolasts in tendon and collagen fibers in group B is weaken than group A, and granulation tissues were growing. The granulation tissues around tendon, the number of cells and collagen fibers in group C became less, he range of the cells was in group C was in disorder, so we can see only a few inflammatory cells infiltrated in group C. Three weeks after operating, the break end in group A was collagen fibers, the cells between sheath and parenchyma less than before, and the capillaries less than before too. The fibrolasts and collagen fibers in parenchyma and sheath of group B were abundant. The break end was linked by collagen fibers and fibroblasts, the range of collagen fibers was little in disorder than group A. A lot of fibroblasts in group C proliferate lively, and the new collagen fibers the range was in disorder, crassitude, inhomogeneous, capillaries proliferate obviously. Four weeks after operation, the tendon break end in group A and B healed both well, the fibroblasts proliferate vigorously differentiation into tenocytes, collagen fibers were well distributed, the range was in regular. Group C is a little worse. The adhesion in group B was the most serious.Six weeks after operation, the quality of healing of group A was the best, the collagen fibers break end were melted in parenchyma of tendon, the border wasn't obvious, capillaries less. The condition of parenchyma in group B was approximate to group A, but the adhesion was more serious; In group C, fibrolasts proliferat vigorously, but tenocytes were less than group A and B clearly, and the differentiation was immature. The collagen fibers proliferate was abundant inhomogeneous, and the arrangement trend to regular but was waves in arrangement. It was abviously that collagen fibers transit from the break end to parenchyma of tendon.The result of image analysis: On the total area of fibroblast and the amount of collagen fiber in most of weeks after operation, there was not statistically significant difference between group A and group B, P>0.05; but between group A,B and group C was statistically significant difference, P<0.05. However, on the total area of fibroblast and the amount of collagen fiber in peri-tendon there was statistically significant difference between group A,C and group B, P<0.05. But between group A and group C wad not statistically significant difference, P>0.05.Conclusion: The method that put the venous indwelling needle in longitudinal proximal near, and put 5.0ng/ml bFGF early in the break end of tendon can promote the process of healing of tendon. It won't promote the level of adhesion between tendon and tissue around the tendon, so that it provides the proof of histology to early stage dirigation. The method is very simple, and the price that the patient can afford is cheap We can apply chemicals conveniently, and may achieve definite results. It provides a new way to recover the tendon damnifi- cation in clinic.