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Withania somnifera is one of the most valuable medicinal plants used in human and veterinary disorders. The present work was proposed to consider the possibility of the antimicrobial effect of in-vivo and in-vitro generated leaf extracts of W. somnifera against some known bacterial strains and the chance of utilizing the constituents present there for the preparation of pharmaceutical products. This is achieved through the zone of inhibition studies and minimum inhibitory concentrations. For this work, four known bacterial strains were used. Among them, two were Gram-positive namely Streptococcus pyrogens and Streptococcus mutans, and two were Gram-negative, namely Salmonella typhimurium and Vibrio cholerae. The inhibition zones of different plant extracts were then compared with standard antibiotics like Neomycin, Kanamycin, and Gentamycin. In comparison to ethanolic extract, methanolic plant extract produces better results. Again, in-vivo generated plant extracts are comparatively better than in-vitro generated plant extracts. The methanolic extract shows maximum antimicrobial activity against Streptococcus mutans, followed by Salmonella typhimurium. For all the plant extracts, Vibrio cholerae shows the least inhibition zone. These opportunities can build up innovative knowledge in the research of medicinal products.

Hydrogel mineralization with calcium phosphate (CaP) and antibacterial activity are desirable for applications in bone regeneration. Mineralization with CaP can be induced using the enzyme alkaline phosphatase (ALP), responsible for CaP formation in bone tissue. Incorporation of polyphenols, plant-derived bactericidal molecules, was hypothesized to provide antibacterial activity and enhance ALP-induced mineralization. Three phenolic rich plant extracts from: (i) green tea, rich in epigallocatechin gallate (EGCG) (herafter referred to as EGCG-rich extract); (ii) pine bark and (iii) rosemary were added to gellan gum (GG) hydrogels and subsequently mineralized using ALP. The phenolic composition of the three extracts used were analyzed by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MSn). EGCG-rich extract showed the highest content of phenolic compounds and promoted the highest CaP formation as corroborated by dry mass percentage meassurements and ICP-OES de-termination of mass of elemental Ca and P. All three extracts alone exhibited antibacterial activity in the following order EGCG-rich > PI > RO, respectively. However, extract-loaded and mineralized GG hydro-gels did not exhibit appreciable antibacterial activity by diffusion test. In conclusion, only the EGCG-rich extract promotes ALP-mediated mineralization.

Polyelectrolyte complexes (PEC) of chitosan and ulvan were fabricated to study alkaline phosphatase (ALP) mediated formation of apatitic minerals. Scaffolds of the PEC were subjected to ALP and successful mineral formation was studied using SEM, Raman and XRD techniques. Investigation of the morphology via SEM shows globular structures of the deposited minerals, which promoted cell attachment, proliferation and extracellular matrix formation. The PEC and their successful calcium phosphate based mineralization offers a greener route of scaffold fabrication towards developing resorbable materials for tissue engineering.

Hydrogels mineralized with calcium phosphate (CaP) are increasingly popular bone regeneration biomaterials. Mineralization can be achieved by phosphatase enzyme incorporation and incubation in calcium glycerophosphate (CaGP). Gellan gum (GG) hydrogels containing the enzyme phytase and chitosan oligomer were mineralized in CaGP solution and characterized with human osteoblast-like MG63 cells and adipose tissue-derived stem cells (ADSC). Phytase induced CaP formation. Chitosan concentration determined mineralization extent and hydrogel mechanical reinforcement. Phytase-induced mineralization promoted MG63 adhesion and proliferation, especially in the presence of chitosan, and was non-toxic to MG63 cells (with and without chitosan). ADSC adhesion and proliferation were poor without mineralization. Chitosan did not affect ADSC osteogenic differentiation.

Mineralization of hydrogels is desirable prior to applications in bone regeneration. CaCO3 is a widely used bone regeneration material and Mg, when used as a component of calcium phosphate biomaterials, has promoted bone-forming cell adhesion and proliferation and bone regeneration. In this study, gellan gum (GG) hydrogels were mineralized with carbonates containing different amounts of calcium (Ca) and magnesium (Mg) by alternate soaking in, firstly, a calcium and/or magnesium ion solution and, secondly, a carbonate ion solution. This alternate soaking cycle was repeated five times. Five different calcium and/or magnesium ion solutions, containing different molar ratios of Ca to Mg ranging from Mg-free to Ca-free were compared. Carbonate mineral formed in all sample groups subjected to the Ca:Mg elemental ratio in the carbonate mineral formed was higher than in the respective mineralizing solution. Mineral formed in the absence of Mg was predominantly CaCO3 in the form of a mixture of calcite and vaterite. Increasing the Mg content in the mineral formed led to the formation of magnesian calcite, decreased the total amount of the mineral formed and its crystallinity. Hydrogel mineralization and increasing Mg content in mineral formed did not obviously improve proliferation of MC3T3-E1 osteoblast-like cells or differentiation after 7 days.

Porous biodegradable scaffolds represent promising candidates for tissue‐engineering applications because of their capability to be preseeded with cells. We report an uncrosslinked chitosan scaffold designed with the aim of inducing and supporting enzyme‐mediated formation of apatite minerals in the absence of osteogenic growth factors. To realize this, natural enzyme alkaline phosphatase (ALP) was incorporated into uncrosslinked chitosan scaffolds. The uncrosslinked chitosan makes available amine and alcohol functionalities to enhance the biomineralization process. The physicochemical findings revealed homogeneous mineralization, with the phase structure of the formed minerals resembling that of apatite at low mineral concentrations, and similar to dicalcium phosphate dihydrate (DCPD) with increasing ALP content. The MC3T3 cell activity clearly showed that the mineralization of the chitosan scaffolds was effective in improving cellular adhesion, proliferation and colonization.

Mineralization of hydrogel biomaterials is considered desirable to improve their suitability as materials for bone regeneration. Calcium carbonate (CaCO3 ) has been successfully applied as a bone regeneration material, but hydrogel-CaCO3 composites have received less attention. Magnesium (Mg) has been used as a component of calcium phosphate biomaterials to stimulate bone-forming cell adhesion and proliferation and bone regeneration in vivo, but its effect as a component of carbonate-based biomaterials remains uninvestigated. In the present study, gellan gum (GG) hydrogels were mineralized enzymatically with CaCO3 , Mg-enriched CaCO3 and magnesium carbonate to generate composite biomaterials for bone regeneration. Hydrogels loaded with the enzyme urease were mineralized by incubation in mineralization media containing urea and different ratios of calcium and magnesium ions. Increasing the magnesium concentration decreased mineral crystallinity. At low magnesium concentrations calcite was formed, while at higher concentrations magnesian calcite was formed. Hydromagnesite (Mg5 (CO3 )4 (OH)2 .4H2 O) formed at high magnesium concentration in the absence of calcium. The amount of mineral formed and compressive strength decreased with increasing magnesium concentration in the mineralization medium. The calcium:magnesium elemental ratio in the mineral formed was higher than in the respective mineralization media. Mineralization of hydrogels with calcite or magnesian calcite promoted adhesion and growth of osteoblast-like cells. Hydrogels mineralized with hydromagnesite displayed higher cytotoxicity. In conclusion, enzymatic mineralization of GG hydrogels with CaCO3 in the form of calcite successfully reinforced hydrogels and promoted osteoblast-like cell adhesion and growth, but magnesium enrichment had no definitive positive effect. Copyright © 2017 John Wiley & Sons, Ltd.

Hydrogels offer several advantages as biomaterials for bone regeneration, including ease of incorporation of soluble substances such as mineralization-promoting enzymes and antibacterial agents. Mineralization with calcium phosphate (CaP) increases bioactivity, while antibacterial activity reduces the risk of infection. Here, gellan gum (GG) hydrogels were enriched with alkaline phosphatase (ALP) and/or Seanol(®), a seaweed extract rich in phlorotannins (brown algae-derived polyphenols), to induce mineralization with CaP and increase antibacterial activity, respectively. The sample groups were unmineralized hydrogels, denoted as GG, GG/ALP, GG/Seanol and GG/Seanol/ALP, and hydrogels incubated in mineralization medium (0.1 M calcium glycerophosphate), denoted as GG/ALP_min, GG/Seanol_min and GG/Seanol/ALP_min. Seanol(®) enhanced mineralization with CaP and also increased compressive modulus. Seanol(®) and ALP interacted in a non-covalent manner. Release of Seanol(®) occurred in a burst phase and was impeded by ALP-mediated mineralization. Groups GG/Seanol and GG/ALP/Seanol exhibited antibacterial activity against methicillin-resistant Staphylococcus aureus. GG/Seanol/ALP_min, but not GG/Seanol_min, retained some antibacterial activity. Eluates taken from groups GG/ALP_min, GG/Seanol_min and GG/ALP/Seanol_min displayed comparable cytotoxicity towards MG-63 osteoblast-like cells. These results suggest that enrichment of hydrogel biomaterials with phlorotannin-rich extracts is a promising strategy to increase mineralizability and antibacterial activity.

Introduction: Mineralization of hydrogel biomaterials is considered desirable to improve their suitability as materials for bone regeneration[1],[2]. Hydrogels have been most commonly mineralized with calcium phosphate (CaP), but hydrogel-CaCO3 composites have received less attention. Magnesium (Mg) has been added to CaP to stimulate cell adhesion and proliferation and bone regeneration in vivo, but its effect as a component of carbonate-based biomaterials remains uninvestigated. In this study, gellan gum (GG) hydrogels were mineralized enzymatically with (CaCO3), Mg-enriched CaCO3 and magnesium carbonate to generate composite biomaterials for bone regeneration. GG is an inexpensive, biotechnologically produced anionic polysaccharide, from which hydrogels for cartilage regeneration have been formed by crosslinking with divalent ions[3]. Methods: GG hydrogels were loaded with the enzyme urease by incubation in 5% (w/v) urease solution and mineralized for 5 days in five different media denoted as UA, UB, UC, UD and UE, which contained urea (0.17 M) and different concentrations of CaCl2 and MgCl2 (270:0, 202.5:67.5, 135:135, 67.5:202.5 and 0:250, respectively (mmol dm-3)). Discs were autoclaved and subjected to physiochemical, mechanical and cell biological characterization. Results: FTIR, SEM, TGA and XRD analysis revealed that increasing magnesium concentration decreased mineral crystallinity. At low magnesium concentrations calcite was formed, while at higher concentrations magnesian calcite was formed. Hydromagnesite formed at high magnesium concentration in the absence of calcium. Amount of mineral formed and compressive strength decreased with increasing magnesium concentration in the mineralization medium. ICP analysis revealed that Ca:Mg elemental ratio in the mineral formed was higher than in the respective mineralization media. Mineralization of hydrogels promoted adhesion and growth of osteoblast-like cells, which were supported best on min

Gene therapy has immense potential to manage the burden of neurological problems. Gene therapy to the central nervous system can be accomplished either by using a viral or a non-viral vector. Non-viral vectors are superior to viral vectors in terms of their safety profiles. However, their significantly low transfection efficiency is a major concern in the development of sustained non-viral gene therapy to the brain. The post-mitotic nature of neurons and the ability of non-viral vectors to cross the blood–brain barrier are the two challenging tasks that have yet to be addressed. Here, in this review, we have summarized the list of cationic polymers used for gene delivery to the brain and associated challenges. Cationic polymers are positively charged molecules that have a ready affinity towards negatively charged DNA/RNA, which allows them to carry genes. Cationic polymers are good candidates for gene therapy in the brain; however, there is scope for improvement to make them better. Development of newer synthetic cationic polymers and modification of natural polymers is the future for brain gene delivery. A biomaterials approach will also help to improve gene therapy. Biomaterial scaffolds can provide gene delivery with a safe degradation profile as well as a depot system that will allow long-term therapeutic effects. Sustained gene therapy in the brain can be achieved by integration of efforts from biology, technology, engineering and nanotechnology.