Sai Rama Krishna Meka

Strontium eluting nanofibers augment stem cell osteogenesis for bone tissue regeneration.

Strontium is known to offer a therapeutic benefit to osteoporotic patients by promoting bone formation. Thus, toward engineering scaffolds for bone tissue regeneration we have prepared polymer nanocomposite scaffolds by electrospinning. Strontium carbonate nanoparticles (nSrCO3) were added to poly(ε-caprolactone) (PCL) at 10 and 20wt% to develop nanocomposite fibrous scaffolds (PCL/SrC10 and PCL/SrC20) with fiber diameter in the range of 300-500nm. Incorporation of nSrCO3 decreased crystallinity and the elastic modulus of PCL. The composite scaffolds released Sr(2+) ions with up to 65ppm in 4days from the PCL/SrC20 scaffolds. Cell studies confirmed that the composite scaffold with 20% nSrCO3 enhanced proliferation of human mesenchymal stem cells in vitro. There was marked increase in mineral deposition up to four folds in PCL/SrC20 suggesting enhanced osteogenesis. This was corroborated by increased mRNA and protein expression of various osteogenic markers such as BMP-2, Osterix and Runx2 in the PCL/SrC20 fibers. Thus, incorporation of nSrCO3 in polymer scaffolds is a promising strategy for bone tissue engineering as an alternative to the use of labile growth factors to impart bioactivity to polymer scaffolds.

Curcumin eluting nanofibers augment osteogenesis toward phytochemical based bone tissue engineering

Curcumin is a phenolic compound isolated from Curcuma longa that is known to exhibit wide ranging biological activity including potential benefits for bone growth. The aim of this work was to engineer curcumin eluting tissue scaffolds and investigate their potential use in bone tissue regeneration. We prepared curcumin loaded poly(ε-caprolactone) (PCL) nanofibers by electrospinning. Morphological characterization of the nanofibers revealed that the average diameter of neat fibers and that of fibers with 1 wt% and 5 wt% curcumin is 840  ±  130 nm, 827  ±  129 nm and 680  ±  110 nm, respectively. Fourier transformation infrared spectroscopy and 1H nuclear magnetic resonance confirmed the successful loading of the drug in fibers. In aqueous medium, the fibers released  ≈18% of the encapsulated drug in 3 d and  ≈60% in 9 d. The cell response to the curcumin loaded nanofibers was assessed using MC3T3-E1 pre-osteoblasts. Cell proliferation was moderated with increased loading of curcumin and was 50% lower on the fibers containing 5% curcumin at day 10 than the control fibers. Osteogenesis was confirmed by assaying the expression of alkaline phosphatase and staining of mineral deposits by Alizarin red stain, which were both markedly higher for 1% curcumin compared to neat polymer but lower for 5% curcumin. Mineral deposition was also confirmed chemically by Fourier transform infrared spectroscopy. These results were corroborated by increased gene and protein expression of known osteogenic markers in 1% curcumin. Thus, controlled release of curcumin from polymer scaffolds is a promising strategy for bone tissue regeneration.

Polyester derived from recycled poly(ethylene terephthalate) waste for regenerative medicine

Despite advances in regenerative medicine, the cost of such therapies is beyond the reach of many patients globally in part due to the use of expensive biomedical polymers. Large volumes of poly(ethylene terephthalate) (PET) in municipal waste is a potential source of low cost polymers. A novel polyester was prepared by a catalyst-free, melt polycondensation reaction of bis(hydroxyethylene) terephthalate derived from PET post-consumer waste with other multi-functional monomers from renewable sources such as citric acid, sebacic acid and D-mannitol. The mechanical properties and degradation rate of the polyester can be tuned by varying the composition and the post-polymerization time. The polyester was found to be elastomeric, showed excellent cytocompatibility in vitro and elicited minimal immune response in vivo. Three-dimensional porous scaffolds facilitated osteogenic differentiation and mineralization. This class of polyester derived from low cost, recycled waste and renewable sources is a promising candidate for use in regenerative medicine.

A self-assembling polycationic nanocarrier that exhibits exceptional gene transfection efficiency

The lack of an efficient and safe carrier is a major impediment in the field of gene therapy. Although gelatin (GT), a naturally derived polymer, is widely used in drug delivery applications, it is unable to bind DNA efficiently. In this study, a novel polycationic gene carrier was prepared by conjugation of low molecular weight polyethyleneimine (LPEI) with GT through 4-bromonaphthaleic anhydride as a coupling agent to avoid self crosslinking. Self-assembly of LPEI conjugated GT (GT–LPEI) with plasmid DNA (pDNA) yielded nanoparticles with high gene complexation ability to form ∼250 nm cylindrical nanoparticles with a zeta potential of ∼27 mV. GT–LPEI showed exceptionally high transfection efficiency (>90%) in various mammalian cells including primary stem cells with minimal cytotoxicity. The transfection efficiency of GT–LPEI significantly surpassed that of many commercial reagents. The high gene transfection expression was confirmed in vivo. Thus, GT–LPEI is shown to be a promising nonviral carrier for potential use in gene therapy.

In situ preparation of multicomponent polymer composite nanofibrous scaffolds with enhanced osteogenic and angiogenic activities

Bioactive ceramics are extensively used for bone repair and regeneration, which release ions to initiate apatite formation and promote osteogenic differentiation eventually resulting in strong bonding to bone. Toward enhancing the bioactivity of polymeric nanofibrous scaffolds, this work presents a one-step in situ sol-gel method to fabricate electrospun composite nanofibrous scaffolds encapsulating well dispersed ceramic nanoparticles overcoming the limitations of current preparation techniques. Transmission electron micrographs revealed uniform distribution of ceramic nanoparticles within the polymer nanofibers. The multicomponent scaffolds were found to release calcium, silicon and phosphate ions that mimic the dissolution and bioactivity of conventional bioactive glasses. The scaffolds enhanced the bioactivity of PCL fibers as observed through enhanced apatite formation in simulated body fluid. The released ions markedly enhanced the proliferation and osteogenic differentiation of human mesenchymal stem cells and the angiogenic activity of human endothelial cells in vitro. This work has important implications for engineering the next-generation nanostructured scaffolds that exhibit multi-biofunctional activities for bone tissue regeneration.

Synthesis of a Block Copolymer Exhibiting Cell-Responsive Phytochemical Release for Cancer Therapy

Phytochemicals constitute a promising class of therapeutics for the treatment of various diseases, but their delivery poses significant challenges. In this work, a nanoscale polyactive emulsion was designed for smart, cell-responsive delivery of a curcumin prodrug (curcumin dicarboxylate, CDA) that was chemically conjugated to enzymatically labile oligo-peptides with polycaprolactone (PCL) as the carrier. Matrix metalloproteinase (MMP)-sensitive (PLGLYAL) or nonsensitive (GPYYPLG) peptides were used as spacers for conjugating CDA and PCL. This CDA nanoemulsion incorporating the MMP-sensitive sequence exhibited markedly higher anti-cancer activity, cell internalization, and generation of reactive oxygen species in cancer cells in vitro than the control with the nonsensitive oligopeptide. Moreover, the nanopolyactives induced minimal cytotoxicity in noncancerous cell line. This work presents a unique strategy to engineer smart nano-polyactives for efficient and targeted delivery of phytochemicals.