2023, Harahap, Iskandar Azmy, Kuligowski, Maciej, Schmidt, Marcin, Kurzawa, Paweł, Suliburska, Joanna

Isoflavones and probiotics are promising nutrients for bone health, and magnesium (Mg) is essential for bone metabolism. This study aimed to determine the effects of daidzein, genistein and Lactobacillus acidophilus on the Mg status of healthy female rats. Forty-eight rats were randomly assigned to six groups, with the control group receiving a standard diet (AIN 93M). The remaining groups were fed the same diet with added ingredients such as tempeh flour; soy flour; pure daidzein and genistein; L. acidophilus or a combination of daidzein, genistein, and L. acidophilus. Tissue samples were collected after the eight-week intervention, and Mg concentrations were analysed using flame atomic absorption spectrometry. Myeloid and erythroid cells were determined using the haematoxylin and eosin bone staining method. Statistical analyses were conducted using one-way ANOVA with Tukey’s post hoc test and Pearson’s correlation coefficient. The threshold for significance was p < 0.05. Compared with the control group, adding tempeh to the diet of rats resulted in significant changes in Mg concentrations in various tissues, with a decrease in the kidneys and an increase in the fur. Although not statistically significant compared to the control group, the tempeh group showed increased Mg concentrations in the femur and spleen. The myeloid-to-erythroid cell ratio did not differ significantly among groups, but all intervention groups showed higher ratios than the control group. A strong negative correlation was observed between Mg concentrations in the kidneys and fur. Conversely, a positive correlation was identified between Mg concentrations in the pancreas and fur. Daily consumption of tempeh may improve Mg status in the organism. Intake of pure daidzein, genistein, or probiotic seems to have no effect on Mg concentrations in healthy rats.

2024, Harahap, Iskandar Azmy, Olejnik, Anna, Kowalska, Katarzyna, Suliburska, Joanna

Adequate calcium intake is crucial for the prevention and treatment of bone-related issues. Developing a nutritional source of readily bioavailable calcium is particularly significant for individuals deficient in this essential element and at risk of developing osteoporosis. This research aimed to evaluate the impact of tempeh (T), daidzein (D), and Lactobacillus acidophilus (LA) within a simulated intestinal environment consisting of Caco-2 epithelial and Saos-2 cells, focusing on their implications for bone mineralization mechanisms. In the initial phase, calcium bioaccessibility from calcium citrate (CaCt), LA, D, the daidzein combination D–CaCt–LA (D1:1:1), and the tempeh combination T–CaCt–LA (T1:1:1) was assessed through digestion simulation. The calcium content of both untreated and digested samples was determined using atomic absorption spectrometry (AAS). In the subsequent stage, the digested samples were used to induce intestinal absorption in differentiated enterocyte-like Caco-2 cells. The permeable fractions were then evaluated in a culture of osteoblast-like Saos-2 cells. Preliminary cellular experiments employed the MTT assay to assess cytotoxicity. The results indicated that the analyzed products did not influence the deposition of extracellular calcium in Saos-2 cells cultured without mineralization stimulators. The combined formulations of permeable fractions of digested CaCt, LA, D, and T demonstrated the capacity to enhance the proliferation of Saos-2 cells. In Saos-2 cells, D, D1:1:1, and LA showed no discernible impact on intracellular calcium accumulation, whereas T and T1:1:1 reduced the calcium deposits. Additionally, mRNA transcripts and alkaline phosphatase (ALP) activity levels in Saos-2 cells cultured without mineralization induction were unaffected by the analyzed products. An examination of the products revealed no discernible effect on ALP activity or mRNA expression during Saos-2 cell differentiation. Our findings suggest that tempeh, daidzein, and L. acidophilus did not positively impact cellular calcium deposition in Saos-2 cells. However, tempeh, daidzein and its combination, and L. acidophilus might enhance the process of osteogenic differentiation in Saos-2 cells. Nevertheless, this study did not identify any synergistic impact on calcium deposition and the process of osteogenic differentiation in Saos-2 cells of isoflavones and probiotics.

2024, Harahap, Iskandar Azmy, Moszak, Małgorzata, Człapka-Matyasik, Magdalena, Skrypnik, Katarzyna, Bogdański, Paweł, Suliburska, Joanna

BackgroundMenopause poses significant health risks for women, particularly an increased vulnerability to fractures associated with osteoporosis. Dietary interventions have emerged as promising strategies, focusing on mitigating the risk of osteoporosis rather than solely addressing the established disease. This 12-week randomized controlled trial aimed to analyze the effects of consuming Lactobacillus acidophilus probiotics on calcium levels, biomarkers of bone metabolism, and bone mineral density (BMD) profiles in postmenopausal women.MethodsFifty-five participants were randomly assigned to receive either a placebo (n = 25) or the probiotic L. acidophilus UALa-01™ (n = 30) daily via oral intervention. Throughout the study, evaluations included body composition, blood biochemical parameters, serum calcium levels, and biomarkers of bone metabolism. Additionally, Dual-energy X-ray absorptiometry was used to measure BMD profiles.ResultsThe findings delineated that the probiotic group experienced a decrease in serum calcium levels compared to their initial levels. However, hair calcium levels and biomarkers related to bone metabolism showed no notable changes within this group. Consumption of probiotic L. acidophilus also seemed to prevent fluctuations in bone turnover markers. Moreover, there were no significant alterations in BMD levels at the lumbar spine, left femur, and total body in the probiotic group. Additionally, probiotic intake led to favorable outcomes by significantly reducing both body fat and visceral fat during the intervention period. Conversely, an adverse effect of consuming probiotic L. acidophilus was observed with a significant increase in glucose concentration.ConclusionIn conclusion, the consumption of L. acidophilus probiotics daily for 12 weeks among postmenopausal women does not affect the profile of BMD, but it may help in stabilizing bone turnover. It is important to note that most measured parameters were within the normal range for this population. However, it is worth noting that 3 months of probiotic supplementation could potentially disrupt calcium and glucose status in postmenopausal women.

2023, Harahap, Iskandar Azmy, Kuligowski, Maciej, Schmidt, Marcin, Suliburska, Joanna

2024, Harahap, Iskandar Azmy, Schmidt, Marcin, Pruszyńska-Oszmałek, Ewa, Sassek, Maciej, Suliburska, Joanna

Osteoporosis in menopausal women requires alternatives to current medications, considering their adverse effects. In this context, probiotics and isoflavone products are promising dietary interventions. The objective of our study was to examine the impacts of Lactobacillus acidophilus and its combination with daidzein and tempeh on calcium status, calcium transporters, and bone metabolism biomarkers in a post-menopausal osteoporotic rat model. A total of 48 female Wistar rats were exposed to a two-stage experiment involving calcium deficit induction and subsequent dietary interventions across six groups. Calcium levels, the gene expression of TRPV5 and TRPV6 calcium transporters, bone histopathology, serum bone metabolism markers, and blood biochemistry were evaluated. The results revealed that, while decreasing serum calcium levels, the groups that received the probiotic L. acidophilus and isoflavone combination exhibited increased bone metabolism biomarkers and decreased calcium transporter expressions, akin to the effects of bisphosphonate. Additionally, significant improvements in bone histopathology were observed in these groups. However, the group receiving probiotic L. acidophilus alone did not exhibit significant changes in bone resorption biomarkers, calcium transporter expression, or various blood parameters. Meanwhile, the combination of probiotic L. acidophilus with tempeh positively influenced hematological parameters and reduced cholesterol and triglyceride levels, but it led to elevated blood glucose levels. Correlation analyses highlighted associations between serum calcium levels, calcium transporter expression, and bone metabolism biomarkers. In conclusion, our findings suggest that the daily consumption of probiotic L. acidophilus in combination with isoflavone products may improve bone health in ovariectomized rats, warranting further research to elucidate potential interactions with other nutrients.

2024, Skrypnik, Katarzyna, Schmidt, Marcin, Olejnik-Schmidt, Agnieszka, Harahap, Iskandar Azmy, Suliburska, Joanna

AbstractBACKGROUNDA high‐fat (HF) diet, diet iron deficiency and iron supplementation may affect inflammatory parameters. Probiotics influence both iron metabolism and inflammation. We compared the inflammatory state in rats on a HF iron‐deficient diet receiving oral iron, Lactobacillus plantarum and Lactobacillus curvatus in different combinations.METHODSThis was a two‐stage experiment. In groups C (n = 8) and HF (n = 8), rats ate a control or HF diet, respectively, for 16 weeks. In the group HFDEF (n = 48), rats ate a HF iron‐deficient diet for 8 weeks (first stage) and were subsequently divided into 6 groups (n = 8 each) receiving the following for a further 8 weeks (second stage): HFDEF – a HF iron‐deficient diet; HFDEFFe – a HF iron‐deficient diet with iron; HFDEFLp and HFDEFLc – a HF iron‐deficient diet with L. plantarum or L. curvatus, respectively; and HFDEFFeLp and HFDEFFeLc – a HF iron‐deficient diet with iron and L. plantarum or L. curvatus, respectively. Body composition analysis and blood sampling was performed. Markers of iron status and levels of total antioxidant status (TAS), C‐reactive protein (CRP), tumour necrosis factor alpha (TNF‐α) and interleukin 6 (IL‐6) were measured in the blood.RESULTSTAS was higher in the HFDEF group (756.57 ± 489.53 ng mL−1) versus the HFDEFLc group (187.04 ± 47.84 ng mL−1; P = 0.022). No more differences were found between groups, or in TAS, CRP, TNF‐α and IL‐6 concentrations. Also, no differences were found between groups for alanine and aspartate aminotransferases, glucose, total cholesterol, low‐ and high‐density lipoproteins and triglycerides. TAS level was positively correlated with ferritin concentration, IL‐6 with TAS and TNF‐α with hepcidin level.CONCLUSIONSSupplementation with L. plantarum, L. curvatus and iron in combinations exerts no influence on inflammatory status, lipid profile, hepatic function and serum fasting glucose in rats on a HF iron‐deficient diet. © 2024 Society of Chemical Industry.

2025, Harahap, Iskandar Azmy, Suliburska, Joanna, Karaca, Asli Can, Capanoglu, Esra, Esatbeyoglu, Tuba

AbstractThe increasing prevalence of metabolic diseases and the global drive toward achieving Sustainable Development Goals (SDGs) underscore the need for sustainable, nutrient‐dense foods. Soybeans (Glycine max), a critical global crop, offer promising solutions; however, their predominant use as animal feed raises concerns regarding food security and environmental sustainability. Fermented soy products—including tempeh, natto, and miso—are rich in bioactive compounds such as peptides and isoflavones, which offer potential therapeutic effects and hold cultural and nutritional significance. These fermented products provide bioactive profiles with unique health‐promoting properties. This review critically examines the bioactive compounds generated through fermentation, focusing on their bioconversion pathways in the gastrointestinal tract and their metabolic implications for human health. Recent consumer demand for novel food ingredients with additional biological benefits has fueled research into advanced extraction techniques, enhancing the functional applications of bioactive compounds from these soy‐based products. This review further explores innovations in extraction methods that improve bioactive yield and sustainability, reinforcing the applicability of these compounds in health‐promoting food interventions. The originality of this review lies in its in‐depth exploration of the gastrointestinal bioconversion of fermented soy bioactive compounds alongside the latest sustainable extraction methods designed to optimize their use. Future research should aim to refine fermentation and extraction processes, investigate synergistic microbial interactions, and develop environmentally sustainable production methods. These efforts have the potential to position fermented soy products as essential contributors to global nutritional security and sustainable food systems, addressing both public health and environmental needs.

2023, Harahap, Iskandar Azmy, Kuligowski, Maciej, Schmidt, Marcin, Kurzawa, Paweł, Pruszyńska-Oszmałek, Ewa, Sassek, Maciej, Suliburska, Joanna

2023, Harahap, Iskandar Azmy, Kuligowski, Maciej, Schmidt, Marcin, Kołodziejski, Paweł, Suliburska, Joanna

AbstractCalcium is essential for maintaining bone health as it contributes to bone formation, remodeling, strength, and density. This study investigated the effect of isoflavones and probiotics on calcium transporters' gene expression, serum calcium levels, and bone metabolism biomarkers in healthy female rats. Forty‐eight female Wistar rats were classified into six groups. Bone metabolism biomarkers (pyridinoline, deoxypyridinoline, parathyroid hormone, and osteocalcin) and serum calcium levels were measured by enzyme‐linked immunosorbent assay (ELISA) and atomic absorption spectroscopy (AAS), respectively. Gene expression of calcium transporters (Trpv5 and Trpv6) was evaluated in duodenum and jejunum tissue samples using quantitative polymerase chain reaction (qPCR). Trpv5 and Trpv6, epithelial calcium channels, play a crucial role in calcium transport and homeostasis in the body. The study consisted of a1‐week adaptation period for the rats to adjust to the controlled conditions, followed by an 8‐week intervention phase. The daidzein and genistein group showed a significant increase in the gene expression of the Trpv6 transporter in the duodenum and a marked decrease in serum pyridinoline levels compared to the control group. The tempeh and soybean groups showed a significant decrease in the gene expression of the Trpv5 calcium transporter in the jejunum. However, no significant influence of the Lactobacillus acidophilus diet on calcium transport and bone metabolism biomarkers was observed in the L. acidophilus group. The correlation analysis showed a significant positive relationship between serum calcium, bone metabolism biomarkers, and calcium transporters. In conclusion, our study demonstrates that the daidzein and genistein diet improves calcium transport in the duodenum and reduces pyridinoline serum concentrations, while tempeh and soybean diets reduce calcium transport in the jejunum. However, the combination of daidzein, genistein, and L. acidophilus did not demonstrate a synergistic effect on calcium transport and bone metabolism, suggesting that further investigations are needed to elucidate their potential interactions.

2025, Harahap, Iskandar Azmy, Salem, Omar, Fahmi, Rifaldi Lutfi, Ahmed, Naglaa, Leciejewska, Natalia, Suliburska, Joanna

Background/Objectives: Postmenopausal osteoporosis disrupts magnesium homeostasis and hematological balance, contributing to systemic inflammation and metabolic alterations. This study aimed to assess the effects of dietary interventions—tempeh, daidzein, probiotics, and their combinations—on magnesium status and hematological ratios in a postmenopausal osteoporotic Wistar rat model. Methods: Sixty-four rats were divided into one sham group (n = 8) and seven ovariectomized (OVX) groups (n = 56), with different modified diets administered for six weeks. In addition, one of the groups received alendronate bisphosphonate as a pharmacological reference to benchmark the dietary interventions against standard anti-osteoporotic therapy. Magnesium levels in the tissues and feces, along with blood hematological ratios (neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), platelet-to-lymphocyte ratio (PLR), and triglyceride-to-glucose index (TyG)), were evaluated. Results: The results revealed that a combination of tempeh and probiotics (OTL) significantly increased magnesium levels in the feces, spleen, and hair, while reducing liver magnesium levels. Compared to the standard groups (S and O), the hematological analysis revealed that the daidzein group (OD) had the highest MLR, while the OTL group exhibited the lowest TyG index. The alendronate bisphosphonate (OB) intervention showed no significant effect on tissue magnesium levels, feces magnesium levels, or hematological ratios. Correlation analysis revealed a strong negative association between spleen magnesium levels and the PLR (r = −0.626) and a positive relationship between liver magnesium levels and TyG (r = 0.422). Conclusions: The authors of this study concludes that while ovariectomy significantly altered magnesium status and hematological ratios, the dietary combination of tempeh, daidzein, and probiotics did not demonstrate an apparent beneficial effect on magnesium status or inflammatory ratios in a postmenopausal osteoporotic rat model. However, the findings highlight interesting aspects of magnesium status and its correlations with metabolic and inflammatory parameters, warranting further investigation.

2024, Harahap, Iskandar Azmy, Kuligowski, Maciej Jakub, Cieślak, Adam, Kołodziejski, Paweł Antoni, Suliburska, Joanna

Menopause marks a critical life stage characterized by hormonal changes that significantly impact bone health, leading to a heightened susceptibility to bone fractures. This research seeks to elucidate the impact of daidzein and tempeh on calcium status, calcium transporters, and bone metabolism in an ovariectomized rat model. Forty female Wistar rats, aged 3 months, participated in a two-phase experiment. The initial phase involved inducing a calcium deficit, while the second phase comprised dietary interventions across five groups: Sham (S) and Ovariectomy (O) with a standard diet, O with bisphosphonate (OB), O with pure daidzein (OD), and O with tempeh (OT). Multiple parameters, encompassing calcium levels, calcium transporters, bone histopathology, and serum bone metabolism markers, were evaluated. The findings revealed that the OT group showcased heightened levels of bone turnover markers, such as pyridinoline, C-telopeptide of type I collagen, bone alkaline phosphatase, and procollagen type I N-terminal propeptide, in contrast to S and O groups, with statistical significance (p < 0.05). Histopathologically, both the OD and OT groups exhibited effects akin to the OB group, indicating a decrease in the surface area occupied by adipocytes in the femoral bone structure, although statistically non-equivalent, supporting the directionally similar trends. Although TRPV5 and TRPV6 mRNA expression levels in the jejunum and duodenum did not display statistically significant differences (p > 0.05), the OD and OT groups exhibited increased expression compared to the O group. We hypothesized that obtained results may be related to the effect of isoflavones on estrogen pathways because of their structurally similar to endogenous estrogen and weak estrogenic properties. In conclusion, the daily consumption of pure daidzein and tempeh could potentially improve and reinstate calcium status, calcium transport, and bone metabolism in ovariectomized rats. Additionally, isoflavone products demonstrate effects similar to bisphosphonate drugs on these parameters in ovariectomized rats.

2023, Harahap, Iskandar Azmy, Suliburska, Joanna