Ipriflavone Group Research Articles

Development of Therapeutic Agent for Osteoarthritis via Inhibition of KIAA1199 Activity: Effect of Ipriflavone In Vivo.

This study aimed to clarify the effects of ipriflavone, which effectively reduces KIAA1199 activity, on osteoarthritis (OA) development and progression in an in vivo OA mouse model. The OA model mice were divided into the ipriflavone (200 mg/kg/day) group and the control group. OA onset and progression were evaluated with the Mankin score, and KIAA1199 expression and hyaluronan (HA) accumulation were analyzed by immunostaining. The molecular weight of HA in the cartilage tissue and serum HA concentration were analyzed by chromatography and competitive HA enzyme-linked immunoassay. The effects of ipriflavone on the bovine cartilage explant culture under the influence of IL-1β were also investigated. In the ipriflavone group, Safranin-O stainability was well-preserved, resulting in significant reduction of the Mankin score (p = 0.027). KIAA1199 staining positivity decreased and HA stainability was preserved in the ipriflavone group. The serum HA concentration decreased, and the molecular weight of HA in the cartilage tissue increased in the ipriflavone group. The results of the cartilage explant culture indicated that ipriflavone could reduce GAG losses and increase the molecular weight of HA. Thus, ipriflavone may have an inhibitory effect on OA development/progression. Ipriflavone could be a therapeutic drug for OA by targeting KIAA1199 activity.

Ipriflavone promotes proliferation and osteogenic differentiation of periodontal ligament cells by activating GPR30/PI3K/AKT signaling pathway.

ObjectivesThis study was performed to investigate the effects and mechanism of ipriflavone (IP) on the proliferation and osteoblastic differentiation of periodontal ligament cells in vitro and periodontal tissue remodeling following orthodontic tooth movement (OTM) in vivo.Materials and methodsHuman periodontal ligament cells (hPDLCs) were cultured in vitro and cell counting kit-8, alkaline phosphatase (ALP) activity assay, plate clone formation assay, and alizarin red staining were used to test proliferation and osteogenic differentiation of hPDLCs. What is more, the expression of ALP, Runx2, and GPR30 was examined by real-time polymerase chain reaction and Western blot. To find out if PI3K/AKT signaling pathway was involved in the process, AKT and p-AKT were examined by Western blot. LY294002 (PI3K signaling pathway inhibitor) and small interfering RNA targeting GPR30 mRNA (siGPR30) were used to verify the function of GPR30-mediated PI3K/AKT pathway in this process. Twenty-four male Wistar rats were randomized into 2 groups, the control group with force application and the IP group with force application plus IP. Morphological changes in the periodontal tissue between roots of teeth were investigated using hematoxylin and eosin (HE) staining and bone morphogenetic protein-2 was detected to assess bone remodeling by immunohistochemical staining.ResultsIn vitro, 10−7 M IP was selected significantly promoting proliferation, ALP activity, colony forming efficiency, and mineral deposition (P<0.05) on hPDLCs. Gene expressions of ALP, Runx2, GPR30, and p-AKT were all upregulated than the control group (P<0.05). According to the mechanism, promotion of ALP and Runx2 interdicted by LY294002 and siGPR30 reduced the activation of PI3K/AKT signaling pathway. In addition, HE staining and immunohistochemical staining results showed that the IP group had more new bone formation in the periodontal tissue compared to the control group in vivo.ConclusionIP can promote the expression of ALP and Runx2 which was probably related to the GPR30-mediated PI3K/AKT signaling pathway. Moreover, IP coordination seemed to have the potential to prevent relapsing following OTM.

Effect of Ipriflavone on Osteoblasts and Osteoclasts during Guided Bone Augmentation

We investigated the effect of ipriflavone (IP) on osteoblasts and osteoclasts during guided bone augmentation (GBA). Each of ten rabbits had two titanium caps placed into its calvarium for GBA. The animals were divided into two groups: No-IP and IP (IP orally, 10 mg/kg daily after GBA). One cap was removed from each rabbit after 3 months, and the remaining cap served as a control. One month after the removal, all of the animals were euthanized, and histologic and histomorphometric analyses were performed. The tissue generated at the test site in the two groups was resorbed, and its original shape and volume were not maintained 1 month after cap removal. In particular, roughly 20% of the newly generated tissue in the No-IP group was resorbed (72.9±2.6% vs. 92.4±3.3%, p<0.05), while approximately 8% was resorbed in the IP group. Furthermore, more osteoblast-like cells were present in the IP group than in the No-IP group (44.4±3.05 vs. 30.2±3.11, p<0.05), and the proportion of osteoclast-like cells was reduced in the IP group compared to the No-IP group (0.59±0.02 vs. 0.76±0.06, p<0.05). Therefore, the amount of mineralized tissue generated appeared to increase as the total IP dose increased. Within the limitations of this experimental model, we conclude that daily intake of IP after GBA inhibits the resorption of the augmented tissue and may be useful for improving the quality of newly generated bone beyond the skeletal envelope.

Effect of ipriflavone on pain due to recent osteoporotic vertebral crush fracture

Abstract Aim: Ipriflavone conserves bone mass in postmenopausal osteoporosis. Salmon calcitonin and alendronate, two other anti‐resorptive drugs, have been found to have analgesic effects in osteoporotic acute vertebral fracture. This study aimed to determine if ipriflavone also possesses such an effect. Methods: Thirty‐two women with recent osteoporotic vertebral fractures were randomly assigned to ipriflavone treatment or placebo groups. Ipriflavone was given at the dose of 200 mg three times a day. Non‐steroidal anti‐inflammatory drugs were given ad libitum in both groups. Calcium carbonate (1 g daily) was administered to all subjects. Intensity of pain at rest, on movement and on pressure, pain rating on a 10‐point visual analogue scale, degree of mobility impairment, and supplementary analgesic were assessed at the end of a 3‐month period in both groups to assess the analgesic effect of ipriflavone. Results: Fourteen subjects in the ipriflavone group and 12 in the placebo group completed the trial. After 3 months, all pain variables had decreased significantly in both groups. Intensity of pain at rest and on pressure and supplementary analgesic use were significantly lower in the ipriflavone group compared to the placebo group. Conclusion: The study shows that ipriflavone has an analgesic adjuvant effect in acute osteoporotic vertebral fracture.

Ipriflavone in the Treatment of Postmenopausal Osteoporosis&lt;SUBTITLE&gt;A Randomized Controlled Trial&lt;/SUBTITLE&gt;

Data on the efficacy and safety of ipriflavone for prevention of postmenopausal bone loss are conflicting. To investigate the effect of oral ipriflavone on prevention of postmenopausal bone loss and to assess the safety profile of long-term treatment with ipriflavone in postmenopausal osteoporotic women. Prospective, randomized, double-blind, placebo-controlled, 4-year study conducted in 4 centers in Belgium, Denmark, and Italy from August 1994 to July 1998. Four hundred seventy-four postmenopausal white women, aged 45 to 75 years, with bone mineral densities (BMDs) of less than 0.86 g/cm(2). Patients were randomly assigned to receive ipriflavone, 200 mg 3 times per day (n = 234), or placebo (n = 240); all received 500 mg/d of calcium. Efficacy measures included spine, hip, and forearm BMD and biochemical markers of bone resorption (urinary hydroxyproline corrected for creatinine and urinary CrossLaps [Osteometer Biotech, Herlev, Denmark] corrected for creatinine), assessed every 6 months. Laboratory safety measures and adverse events were recorded every 3 months. Based on intent-to-treat analysis, after 36 months of treatment, the annual percentage change from baseline in BMD of the lumbar spine for ipriflavone vs placebo (0.1% [95% confidence interval (CI), -7.9% to 8.1%] vs 0.8% [95% CI, -9.1% to 10.7%]; P =.14), or in any of the other sites measured, did not differ significantly between groups. The response in biochemical markers was also similar between groups (eg, for hydroxyproline corrected for creatinine, 20.13 mg/g [95% CI, 18.85-21.41 mg/g] vs 20.67 mg/g [95% CI, 19.41-21.92 mg/g]; P =.96); urinary CrossLaps corrected for creatinine, 268 mg/mol (95% CI, 249-288 mg/mol) vs 268 mg/mol (95% CI, 254-282 mg/mol); P =.81. The number of women with new vertebral fracture was identical or nearly so in the 2 groups at all time points. Lymphocyte concentrations decreased significantly (500/microL (0.5 x 10(9)/L]) in women treated with ipriflavone. Thirty-one women (13.2%) in the ipriflavone group developed subclinical lymphocytopenia, of whom 29 developed it during ipriflavone treatment. Of these, 15 (52%) of 29 had recovered spontaneously by 1 year and 22 (81%) of 29 by 2 years. Our data indicate that ipriflavone does not prevent bone loss or affect biochemical markers of bone metabolism. Additionally, ipriflavone induces lymphocytopenia in a significant number of women.

Comparison of antiresorptive activities of ipriflavone, an isoflavone derivative, and elcatonin, an eel carbocalcitonin

Thirty postmenopausal women with reduced bone mineral density were divided randomly into two groups based on the chronological sequence of their first visit to the Osteoporosis Clinic of Katsuragi Hospital. Group I was given 600 mg ipriflavone orally daily and group II was weekly injected intramuscularly with 20 units elcatonin, Asu1-7 eel calcitonin (carbocalcitonin). Lumbar spine BMD was measured by dual-energy X-ray absorptiometry, and trabecular bone mineral density at the distal radius, cortical bone density, and relative cortical volume at the radial diaphysis by peripheral computed tomography before the beginning of the study and at the 4th, 8th, and 12th month. Markers of bone metabolism--serum total alkaline phosphatase, bone-specific alkaline phosphatase, tartrate-resistant acid phosphatase, osteocalcin, intact osteocalcin, PICP and ICTP, and urinary pyridinoline, deoxypyridinoline, and calcium/creatinine (Ca/Cr)--were also measured at the same interval. Plasma parathyroid hormone (PTH) and calcitonin (CT) were measured at the same time. Radial trabecular bone density showed a significantly higher rate of increase in group I (ipriflavone group) than in group II (elcatonin group) at the 4th month, whereas lumbar spine BMD showed more pronounced increase in the elcatonin group than in the ipriflavone group throughout the study period. Bone metabolism markers tended to decline in both groups. Total and intact osteocalcin showed a significant fall from the baseline throughout the study period only in the ipriflavone group. Urine pyridinoline and deoxypyridinoline showed a significant fall from the baseline at the 12th month only in the ipriflavone group. On comparing bone gainers with increase of lumbar spine BMD by 2% or more with bone losers with a decrease by 2% or more, only urine Ca/Cr was significantly different, lower in the former than in the latter, despite the general tendency for bone resorption markers to decrease in bone gainers and to increase in bone losers.

Effect of Ipriflavone???a Synthetic Derivative of Natural Isoflavones???on Bone Mass Loss in the Early Years After Menopause

We studied whether oral administration of ipriflavone, a synthetic derivative of naturally occurring isoflavones, could prevent bone loss occurring shortly after menopause. Fifty-six women with low vertebral bone density and with postmenopausal age less than five years were randomly allocated to receive either ipriflavone, 200 mg three times daily, or placebo. All subjects also received 1,000 mg elemental calcium daily. Vertebral bone density declined after two years in women taking only calcium (4.9 +/- 1.1%, SEM, p = 0.001), but it did not change in those receiving ipriflavone (-0.4 +/- 1.1%, n.s.). A significant (p = 0.010) between-treatment difference was evidenced at both year 1 and year 2. At the end of the study, urine hydroxyproline/creatinine excretion was higher in the control group than in the ipriflavone group, as compared to no difference at baseline. Five patients taking ipriflavone and five taking placebo experienced gastrointestinal discomfort or other adverse reactions, but only one and four subjects, respectively, had to discontinue the study. Ipriflavone prevents the rapid bone loss following early menopause. This effect is associated with a reduction of bone turnover rate.

Bone density changes in postmenopausal women with the administration of ipriflavone alone or in association with low-dose ERT

Ipriflavone is a synthetic flavonoid that has been shown to exert a direct inhibitory effect on osteoclastic activity and possibly stimulate the osteoblast activity in different experimental models. The aim of the present study was to evaluate the effects of either ipriflavone alone or ipriflavone plus low dose hormone replacement therapy (HRT) in the prevention of postmenopausal bone loss. Patients were randomly allocated to different treatment groups receiving calcium (500 mg/day, control group), continuous HRT (25 or 50 μg/day of transdermal 17β-estradiol) plus medrogestone 5 mg/day for 12 days/month, ipriflavone at the standard dose of 600 mg/day, or finally ipriflavone 600 mg/day plus 17β-estradiol 25 μ g/day plus medrogestone 5 mg/day for 12 days/month. No significant differences in basal levels of biochemical markers of bone turnover or in basal bone mineral density (BMD) values were evident in the different groups. In the control group after 12 months, spine BMD showed a significant (p < 0.05) 3.41% decrease. The pattern of BMD modification was significantly different from controls in the high dose HRT group (+ 1.84%), the ipriflavone group (+ 0.11%), and the combined ipriflavone/HRT group (-0.22%). Conversely, the BMD pattern in the low dose HRTgroup (-0.55%) was similar to that observed in controls. Thus, present results confirm that ipriflavone and 50 μg/day of transdermal 17β-estradiol are effective measures in the prevention of postmenopausal osteopenia. A lower transdermal estrogen dose is unable to increase the antiresorptive effect of ipriflavone and did not exert any further action in the prevention of postmenopausal osteopenia.

Metabolic and bone effects after administration of ipriflavone and salmon calcitonin in postmenopausal osteoporosis

Forty postmenopausal women with bone mineral density (BMD) > 2 standard deviations below the mean value for healthy age matched controls were enrolled into an open controlled study to evaluate the metabolic and bone effects of ipriflavone (IP) versus salmon calcitonin (sCT) over a 12 month period. Both treatments significantly increased BMD after 6 and 12 months. A 4.3% increase of BMD was obtained in the IP treated group and a 1.9% in the sCT treated group after 12 months (p < 0.001 between treatments). Bone metabolism markers (serum osteocalcin, alkaline phosphatase, urinary calcium and hydroxyproline/creatinine ratio) were significantly reduced in both groups (p < 0.001). The reduction of urinary hydroxyproline/creatinine ratio was significantly greater (p < 0.05) in the IP group after 12 months. Both treatments were well tolerated. Four patients in the IP group reported gastralgia while two patients in the sCT group reported pruritus and one patient epistaxis.

Ipriflavone-treatment of senile osteoporosis: results of a multicenter, double-blind clinical trial of 2 years

Eighty-four out of 100 enrolled female patients affected by osteoporosis completed a double-blind, randomized trial, using ipriflavone (IP) in a dose of 3 × 200 mg/day (41 patients) or placebo (P)(43 patients). All patients received 1g/day oral calcium supplementation. Inclusion criteria were: age over 65 years; at least one vertebral fracture in the past; bone mineral density measured at the distal tenth of the radius lower than the normal average −2 × S.D. The trial period was between June 1990 and November 1993. Patients of the IP group showed a significant increase in bone mineral density ( P < 0.05) compared with the starting values during the whole study period. Pain decreased rapidly, intake of analgesics dropped, and often significant decreases in calciuria, hydroxyprolinuria, alkaline phospatase, osteocalcin and parathormone values were observed. Only two new fractures occurred during the trial in the IP group. Patients of the P group at the end of the study displayed decreased bone mineral densities ( P < 0.05), increased pain, greater consumption of analgesics, and often significant increases in the bone metabolism parameters listed for the IP group. In the

Effect of ipriflavone on bone mineral density of lumbar spine of aged women

Aged women with osteoporosis were followed for 6 months with or without ipriflavone (IP) 600 mg/day, and lumbar bone mineral density (BMD) and bone mineral content (BMC) were evaluated by dual-photon absorptiometry. Although a significant reduction of lumbar BMD was observed in the control group at the end of the experiment, the group treated with IP showed insignificant change. The frequency of deterioration was lower in the IP group compared to the control group. Additionally, the IP effect was more apparent in patients with lower calcitonin and higher bone gla-protein (BGP) levels. These results suggest that IP effectively maintains BMD and BMC of lumbar bone among aged women with osteoporosis, and its activity may be enhanced in patients with low calcitonin and high BGP levels.

Stimulatory effect of ipriflavone on formation of bone-like tissue in rat bone marrow stromal cell culture.

The effects of ipriflavone (IP) (10(-5) M) on bone formation were studied in stromal cells from the femoral bone marrow of young adult rats cultured for 21 days in the presence of beta-glycerophosphate and dexamethasone. Stereoscopic microscopy showed nodule formation after 14 days of culturing, and both the number and the size of the nodules increased with time. The alizarin-red-stained calcified area in the nodules in the IP group was nearly 4 times as large as that in the control after 21 days. Light and electron microscopy revealed the presence of many osteoblast-like cells with developed rough endoplasmic reticulum and Golgi apparatus in the nodules in the control group after 14 days, and a collagenous fibril network was seen among the cells. After 21 days, calcification of the dense collagenous fibril network and bone matrix-like tissue were observed in many nodules, resulting in the formation of bone-like tissue containing osteocyte-like cells. In the IP group, the collagenous fibril network area in the nodules was greater than that in the control after 14 days, and a further increase in both the dense collagenous fibril network area and calcified bone-like tissue area was observed after 21 days. These findings indicate that IP stimulates bone-like tissue formation in the rat bone marrow stromal cell culture, suggesting that the promotion of collagen production by osteoblasts is involved in the stimulation of bone-like tissue formation by IP.