Study Design and Selection of Patients
The study was placebo controlled, randomized, double blind with four groups of subjects. Patient recruitment took place from March 10, 2009 to August 30, 2009 at the second Medical Clinic, Craiova Emergency Hospital, Romania, from the surrounding community. At the screening visit, after providing written informed consent, each participant underwent a screening interview. Based on the screening, subjects were declared eligible to proceed if all inclusion criteria were fulfilled and no exclusion criteria were met. This single-center trial was approved by the Institutional Ethics Committee of the University of Medicine and Pharmacy from Craiova, Romania, according to decision no. 364 from March 2008. The trial is also in agreement with the Helsinki Declaration from 1975, which was revised in 1983.
The number of total enrolled patients was 116 (see Fig. 1). Not all enrolled subjects were included in the intent-to-treat (ITT) population. This ITT population was defined as being all the subjects who received the product and who had some follow-up evaluation. Forty-four people did not meet inclusion criteria or refused to participate were not taken into consideration for the ITT analysis. Thus, the total number of subjects that were included in the ITT analysis was 72. These 72 patients were divided into four groups (19 in group 1, 18 in group 2, 17 in group 3, and 18 in the placebo group), but 12 of them did not meet the entire protocol.
From the total number of patients included in the study (n = 116), only 60 completed the 2-week treatment period. Age span was between 59 and 68 years, and the number of women was higher than the number of men (female, 43 (71.66%) vs. male, 17 (28.33%)). There were 51 rural patients and nine urban patients. The boron dietary intake was calculated to be between 1.9 and 2.2 mg boron per day, uniformly distributed between genders. The number of patients with dyslipidemia and the number of those with nonspecific inflammation was heterogeneous between groups (ranging from 66% for group 2 to 93% for group 1 and 53% for group 2 to 93% for the placebo group, respectively). The baseline demographic characteristics of patients that successfully completed the study are given in Table 1.
Participants Inclusion Criteria
Men and nonpregnant women, between 40 and 85 years old, with primary OA of at least one knee were included. Primary OA was defined by the deterioration and abrasion of the articular cartilage (joint space narrowing) or by the formation of a new bone (osteophytes) at the knee joint surface (medial tibiofemoral, lateral tibiofemoral, or patellofemoral). This fact was demonstrated upon radiological examination that was performed within the previous 3 months.
Participants Exclusion Criteria
Subjects with digestion problems, with fever and/or under treatment with antibiotics, with fructose intolerance and those taking any painkillers and/or vitamin B6 and aspirin were excluded from the study.
Study Visits and Administration
The study used a commercial dietary supplement FruiteX-B® trademark, registered by the FutureCeuticals Company from the USA. FruiteX-B® was scientifically proven as being identical to naturally occurring plant-based boron carbohydrate, CF .
The four subjects groups are described as follows:
Group 1: 28.5 mg CF (1.5 mg boron/day) twice per day
Group 2: 56.5 mg CF (3.0 mg boron/day) twice per day
Group 3: 113 mg CF (6.0 mg boron/day) twice per day
Placebo group: 120 mg placebo twice per day. Placebo material was based only on fructose
The duration of the treatment was 2 weeks. Two capsules were administered per day and were taken orally with meals. The subjects’ interviewers carried out a survey of dietary intake. They presented tableware and food models and investigated the meal intake on two different weekdays and one weekend day, relying on the recall method. Nutrient intake was calculated by the DietSYS+Plus usage (version 5.9) and dietary analysis program (Block Dietary Data Systems). The DietSYS+Plus database, a software that analyzes nutrients, was expanded for the present study to include dietary boron values in the Romanian consumed foods. After computing nutrients intake per subject, intake percentage was calculated in relation to the Dietary Reference Intakes for Romania [30, 31]. Subsequently, using the boron content database of the foods commonly consumed by the Romanian urban and rural people, boron intake was calculated. The computation was based on an analytical boron nutrient database that was created in Romania and previously developed for estimating boron intake (Scorei et al., unpublished results).
Outcomes Measures (Biochemical Parameters)
For biochemical analyses, venous blood samples were taken in the morning after fasting both at the beginning of the study and after 2 weeks of treatment. Commercially available collection tubes without anticoagulant were used to collect blood for the determination of biochemical parameters. Basic biochemical parameters, such as lipid profile (total, HDL, and LDL cholesterol) and inflammatory markers (CRP) were analyzed in serum using standard biochemical procedures. For the inflammatory markers ESR and FBR, anticoagulant was used. Enzymatic colorimetry methods using commercial kits (Boehringer Mannheim, Germany) and a Hitachi 911 automatic chemistry analyzer  measured cholesterol and triglyceride. Assays for CRP were performed using Tina-quant® immunoturbidimetric kit for CRP also available from Boehringer Mannheim Diagnostics, and the applications were developed for the BM/Hitachi 911 . The ESR was determined by the Westergren method using anticoagulant (ethylenediaminetetraacetate)-containing whole blood . The ratio between blood and anticoagulant was 4:1. The reading is done on the integral blood at room temperature after at most 2 h after blood donation. FBR levels were determined in citrated plasma using a kinetic method, as described by Hemker et al. . The blood donation was realized a jeun (on fasting—12 h of post) from venous blood in vacutainers. The serum was separated by centrifugation.
Three emergency telephone numbers were given to the subjects to maintain contact during the study in case they had any adverse events or other concerns related to the study. Each subject was interviewed during site visits to solicit information on possible adverse effects they might have encountered. Participants were instructed to inform the test supervisor if they chose to discontinue the study due to adverse effects.
The CF or placebo effect was evaluated with two-way randomized block analysis of variance. Microsoft Excel software was used for the statistical analysis. Statistical significance was defined at the level of 95% (p < 0.05). Results are expressed as means ± SD.