Subjects

 

Group 1

Enrolled into this study were 15 patients (10 females, five males) with Blount’s disease with a mean age of 3.8 ± 1.1 (SD), range two-to-five years and mean body weight 14.0 ± 2.4 kg. These children were attending the outpatient orthopedic clinic of the University College Hospital, Ibadan. The diagnosis of Blount’s disease was made in these subjects on the basis of clinical features and standard radiographic parameters which were: physiologically bow legs, tibia torsion, medial beaking of the proximal tibia metaphysis on plain radiographs, and sometimes laxity of the lateral collateral ligaments, which was proportionate to the degree of the varus deformity. The radiographs of the patients revealed that they were all between Langenskiold stages III and V, and none was in stage VI.

Group 2

Fifteen (nine females, six males) apparently normal subjects within the same age range as the patients with Blount’s disease who were attending the surgical outpatient clinic for minor surgical elective complaints, such as inguinal hernia and undescended testis, served as controls. The mean body weight was 14.3 ± 2.3 kg (range 10-17 kg). All these patients had normal knees, and none of the subjects was suffering from any metabolic bone disease or malnutrition or on a special diet or medication that may interfere with bone metabolism. viagra soft

Informed consent was obtained from the parents of the children, and ethical approval of the study was obtained from the University College Hospital/College of Medicine, University of Ibadan Ethical Board before the commencement of the study.

MATERIALS

Blood Samples

Ten ml of venous blood was collected from each patient. The samples were allowed to clot and retract for 30 minutes. Blood samples were centrifuged at a relative centrifugal force of 3,000 xg for 10 minutes, and serum was immediately separated from the cells and stored at -20°C, until analyses were carried out.

LABORATORY MEASUREMENTS

Methods

For each assay, internal quality control samples were always included in the assay system and results were only accepted when values fell within the expected range (x ± 2SD). Serum alkaline phosphatase activity was assayed using the method of Anon. The procedure is based on the hydrolysis of p-nitrophenyl phosphate to p-nitrophenol, which produces a yellow color. The color intensity was measured at an absorbancy of 660 nm. canadian pharmacy cialis

The method of Trinder was employed in the estimation of total serum calcium. The process involved precipitation using naphthahydroxamic acid. The precipitate was suspended in alkaline EDTA and heated in a boiling water bath to complete dissolution. Further reaction with ferric nitrate produced a red color, which was measured at an absorbancy of450 nm.

The modified method of Delsal and Manhouri was employed in the estimation of inorganic phosphates in the sera. The method is a modified form of the phosphomolybdic acid reaction. Acetate buffer controlled the pH and paramethylaminophenol sulfate (Rhodol) was employed as the reducing agent. The blue-colored complex was stable for at least 30 minutes and measured at an absorbancy of 880 nm.

Serum zinc was determined by the method of Smith et al., while serum copper was determined by the method of Robinson. Atomic absorption spectrophotometer was used for the determination of these two micronutrients using Pye Unicam S.R 90A series 2 model, New York.

Statistical Analysis

The means and standard deviations (SD) were calculated using a statistical computer package (SPSSVPC). Pairwise comparisons were made using Wilcoxon Two Sample Test, and p<0.05 was regarded as significant.