| BackgroundClubfoot, with reported incidence of1‰to6‰,is a deformity which occursmost frequently in left-foot in children and sometimes in two feet with the incidenceof40%to50%, including Idiopathic Clubfoot (ICF) and Clubfoot which Secondaryto Tethered Cord Syndrome(TCS) more. the occurrence proportion of male andfemale of Clubfoot is3:1, leading foot disability and Impairment of ambulatoryfunction. Moreover, the deformity caused by Clubfoot impact the function of forefoot,mid-foot, hind-foot, ankle and calf. However, the mechanisms of Clubfoot have beenunknown, the foot joints motion in which is limited seriously, especially in ClubfootSecondary to TCS, and children with clubfoot will lead to problems with standing andwalking. Numbers of researchers have investigated the pathogen of Clubfoot in allmanner of ways，the argue of which focus on bones and muscular tissue of the legsand feet. There are many investigated evidences that muscle atrophy especially ingastrocnemius occurs in the patients with Clubfoot.ObjectiveTo investigate the changes of ultrastructure of gastrocnemius in ICF andClubfoot secondary to TCS by scanning electron microscope and transmission electron microscopy, so that we can make it clear that the ultrastructure ofgastrocnemius changes in gastrocnemius in Clubfoot in order to offer further supportfor the theory of proper selection of operation ways and the reduce of postoperativerecurrence rate.Methods1．Material24cases of ICF and24case of Clubfoot secondary to TCS in patients receivedsurgical treatment from August2011to August2013,32feet of which are Clubfootsecondary to TCS and the rest are ICF，and the age of patients is from2to7，theaverage of which is4.7. Among all these cases,15male case and9female cases werediagnosed with ICF in which the number of Talipes Equinovarus occurred in onefoot and Feet is16and8.while the others were Talipes Equinovarus Caused byTethered Cord Syndrome the number of which is17in male and7in female with18cases in one foot and6cases in feet. Distal Muscle tissue were sampled during theoperation, and analyzed by transmission electron microscope. The entire musclesample with strip-type removed from2cm bellow muscle belly in junction of musclebelly and tendon, along the muscle fiber direction, removing the extruding andpulling tissue, and the shape of which is1mm×1mm×2mm.All of the case sampled above with informed consents had authorization byhospital ethics committee.2．MethodsSample fixing: formaldehyde solution, the coagulation of which is10%, wasused to fix the muscle under microscopy; muscle samples observed by electronicmicroscopy were fixed by glutaraldehyde after removing, all of samples are stored at4℃。 Results1. Light microscopy of gastrocnemius in ICF and Secondary ClubfootThe results of light microscopic observations: There were columnar musclefibers in both of the group with the oval cell nucleus on the inside of sarcolemma andlots of myofibril in sarcoplasm which was filamentous with cross striations and visualIband and Aband. However, in group of Clubfoot secondary to TCS, there wereserious vacuolization, fibrosis and fatty changes in myofilament of gastrocnemius;some of the myocyte became apoptosis and funicular nuclei and a mass of fibroblastcould be observed. On the contrary, there existed just Minutes or alterations in ICF.2.electronic microscopy of gastrocnemius in ICF and Secondary ClubfootICF: muscle bundle ranked neatly; muscle lost, abnormal morphology andatrophy existed, collagenous fiber could be observed marked enlargement,nevertheless parts of muscle cells is normalClubfoot secondary to TCS: There were marked enlargements among musclebundles and cellular edema, moreover the sarcomere was not only disorganized andbroadening but the morphology of nuclei of various sizes could also be viewed.Besides that, myofilament tonic contraction existed in Clubfoot secondary to TCSwith parts of collagenous fiber became enlarge.3.Results of the observation of gastrocnemius muscle with transmission electronmicroscope in patients with idiopathic and secondary clubfootThe observation in the ICF group: The arrangement of myofilament disorderedmildly, the Z line widened slightly, and H band shortened mildly. The cells increasedin size, and had abundant cytoplasm. Endoplasmic had reticulum, mitochondriaswelled, and its partial cristae and a bit part of membrane had fusion, and blur, andpartial cristae had dissolution, fracture, and loss. The counts of Glycogen decreasedslightly, and some sarcoplasmic had mild reticulum. The longitudinal, collagenousfibers had periodic stripes and coarse fibers, and arranged in neat rows. In Cross section, fibers were circular or oval, whose diameter increased, and arrangedcompactly, The nucleus was big and clear, the myostroma in the end of nucleus undercristae film reduced, mitochondria had cavitation, and cristae disordered.The observation in the TCS group with secondary talipes equinovarus: partialcristae and membrane of the mitochondria had fusion and blur, and much more cristaehad dissolution, fracture. and loss.Much more mitochondria had vacuolation, atrophy,rupture and reduction, whose vacuolization was more severe. The Sarcomere wasarranged in disorder. The matrix loosed, the gap of muscle myofibrils enlarged, andits number reduced. The number of the mitochondria and the glycogen among musclefibers decreased. The myofilament swollen, the sarcoplasm lost, and the myofilamenthad tonic contraction, The Myofibrillar structure was unclear, and arranged indisorder, Z line and H line were in disorder and unclear, and had poor malalignment,incompleteness and dissolution. The myofibrillar cytoplasm had degeneration andnecrosis, and its membrane disappeared, only to see the bare nucleus.ConclusionsPathological changes such as myofilament arranged, nucleus pycnosis andprogressive atrophy in gastrocnemius substantially contribute to the pathogenesis ofICF and ICF caused by Tethered Cord Syndrome, especially the latter, thePathological changes above is one of the reasons why the more serious talipes exist. |
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