Biosynthesis of Nano-Calcite and Nano-Hydroxyapatite by the Probiotic Bacteria of Bacillus subtilis and Bacillus coagulans

Background and objective: In recent years, the green synthesis of nanomaterials has received more attention than chemical synthesis due to its eco-friendly and compatibility. Material and methods: In this study Bacillus subtilis and Bacillus coagulans, two potential probiotics, were used and grown Bacillus in medium culture containing insoluble calcium phosphate produced nano-hydroxyapatite and nano-calcite. The nano-hydroxyapatite was purified from nano-calcite by heat treatment and washing with a 200 nm filter. The structures, characteristics, and elemental analysis of nano-sized material were studied by Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscope, ultraviolet-visible spectroscopy, energy dispersive X-ray, and X-ray fluorescence. Results: The results showed that hydroxyapatite is made only in a medium containing insoluble calcium phosphate sublimated with urea which is induced phosphatase and urease. Here, for the first time, the braided bacterial nano-hydroxyapatite similar to the bone structure was made in the medium, which caused the production of urease and phosphatase (Maximum 99 U/L) enzymes, and the particle size was less than 100 nm. The ratio of calcium to phosphorus in crude hydroxyapatite and calcite crystal particles made by B. coagulans was 2.9, however, this ratio for pure hydroxyapatite was 1.7. Conclusion: Since the particles are made by antibacterial probiotics, the biological production of these particles makes them a suitable candidate to be used in food, toothpaste, and sanitation products. Braided hydroxyapatite can substitute the needle-like type of food additives for infants and elders due to its safety

Osteoblastic Differentiation of Stem Cells from Human Exfoliated Deciduous Teeth by Probiotic Hydroxyapatite

Objective: Multipotent cells derived from human exfoliated deciduous teeth (SHED) possess the ability to differentiateinto various cell types, including osteoblasts. This study aims to simulate the growth induction and osteogenicdifferentiation of SHED cells using probiotics and their resultant biomaterials.Materials and Methods: This experimental study proceeded in two stages. Initially, we evaluated the effect ofautoclaved nutrient agar (NA) grown probiotic Bacillus coagulans (B. coagulans) on the SHED and MG-63 cell lines.Subsequently, probiotics grown on the Pikovskaya plus urea (PVKU) medium and their synthesised hydroxyapatite (HA)were identified using X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray (EDX), andFourier transform infrared spectroscopy (FTIR), and then used to stimulate growth and osteogenic differentiation of theSHED cell line. Osteoblast cell differentiation was assessed by morphological changes, the alkaline phosphatase (ALP)assay, and alizarin red staining.Results: There was a substantial increase in SHED cell growth of about 14 and 33% due to probiotics grown on NAand PVKU medium, respectively. The PVKU grown probiotics enhanced growth and induced stem cell differentiationdue to HA content. Evidence of this differentiation was seen in the morphological shift from spindle to osteocyte-shapedcells after five days of incubation, an increase in ALP level over 21 days, and detection of intracellular calcium depositsthrough alizarin red staining-all indicative of osteoblast cell development.Conclusion: The osteogenic differentiation process in stem cells, improved by the nano-HA-containing byproducts ofprobiotic bacteria in the PVKU medium, represents a promising pathway for leveraging beneficial bacteria and theirsynthesised biomaterials in tissue engineering

The Therapeutic Effects of a Medicinal Plant Mixture in Capsule Form on Catalase Levels in the Semen of Men with Oligospermia

Objective: In the present study, the therapeutic effects of mixed herbs (onion, ginger, basil, cinnamon, orange peel, yellow and red watermelon seeds, and carrot seed) on catalase levels in the semen of men with oligospermia were evaluated. About 50% of recognized infertility factors are male-related factors, and are mainly the result of oligospermia, astenospermia, and teratozoospermia. Materials and Methods: The study participants included 40 males with oligospermia and infertility. The studied medicine were 700 mg capsules containing onion, ginger, basil, cinnamon, orange peel, yellow and red watermelon seeds, and carrot seed (100 mg of each). Catalase activity was measured by Aebi method. Results: A significant increase was observed in catalase level in semen as a result of using the medicinal plant mixture (P < 0.001). Conclusion: Free radicals play an important role in male infertility. Antioxidants can prevent the damaging effects they have on sperm. Oxidative stress reduction can increase the chances of natural fertility or assisted reproductive technology (ART). Medicinal plants have low costs, complications, and easy availability, and cause an increase in semen plasma antioxidants and subsequent improvement in semen parameters. Thus, they can be the source of new hopes for the treatment of infertility