Uma Tiruchirapalli Sivagnanam

Fabrication and characterization of a collagen coated electrospun poly(3-hydroxybutyric acid)–gelatin nanofibrous scaffold as a soft bio-mimetic material for skin tissue engineering applications

Wound healing is a global health care problem. The use of a suitable dressing material by means of a nanofibrous scaffold with traditionally important medicine can help to repair the damaged skin tissue. An ideal wound dressing material should mimic the function of an extracellular matrix with its improved physiochemical, biological and antimicrobial properties. In this study, the significance features of a collagen coated electrospun poly(3-hydroxybutyric acid)–gelatin nanofibrous scaffold with a bioactive Coccinia grandis extract (CPE) meets the requirements for a wound dressing material. The nanofibrous scaffold with collagen has an attraction for fibroblast, which increases cell adhesion and proliferation. The fabricated nanofibrous scaffold with collagen was characterized physio-chemically using Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and it showed acceptable antibacterial property with both Gram positive and Gram negative bacteria. The thermal and in vitro stability of the nanofibrous scaffold was studied and it was found to have stability more than that required for a wound dressing material. The nanofibrous scaffold supports good swelling property with better porosity for oxygen permeability. The mechanical property of the nanofibrous scaffold showed a Youngs modulus of 2.99 ± 0.16 MPa. The biocompatibility of the nanofibrous scaffold exhibits increased cell adhesion and proliferation of both NIH 3T3 fibroblast and human keratinocytes (HaCaT) cell line. The in vitro fluorescence staining of the nanofibrous matrix using Calcein AM and DAPI exhibits the cell material interaction of the collagen coated nanofibrous scaffold corresponding to increased cell adhesion and proliferation. This approach with a nanofibrous scaffold coated with collagen can be a promising tool in skin tissue engineering and can be useful as a wound dressing material in skin tissue engineering applications.

Design and characterization of 3D hybrid collagen matrixes as a dermal substitute in skin tissue engineering

The highly interconnected porous dressing material was fabricated with the utilization of novel collagen (COL-SPG) for the efficient healing of the wound. Herein, we report the fabrication of 3D collagen impregnated with bioactive extract (COL-SPG-CPE) to get rid of infection at the wound site. The resultant 3D collagen matrix was characterized physiochemically using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and mechanical property. The dressing substrate possesses the high swelling ability, increase in the porosity, in vitro enzymatic degradability and antibacterial property. The in vitro biocompatibility and fluorescence activity of the collagen scaffold against both NIH 3T3 fibroblast and Human keratinocyte (HaCaT) cell lines assisted in excellent cell adhesion and proliferation over the collagen matrix. Furthermore, the in vivo evaluation of the COL-SPG-CPE 3D sponge exhibited with enhanced collagen synthesis and aids in faster reepithelialization. However, the rate of wound healing was influenced by the expression of vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and transforming growth factor (TGF-β) growth factors promotes the collagen synthesis, thereby increases the healing efficiency. Based on the results, COL-SPG-CPE has a potential ability in the remodeling of the wound with the 3D collagen as wound dressing material.

Durable keratin-based bilayered electrospun mats for wound closure

A bilayered nanofibrous scaffold with rapid wound healing properties is found to be suitable for tissue regeneration applications. The objective of this study is to reveal the fabrication of a poly(3-hydroxybutyric acid) (P)-gelatin (G) nanofibrous mat through electrospinning, with a horn keratin-chitosan-based biosheet (KC) as a bilayered nanofibrous scaffold. The mupirocin (D)-loaded horn KC biosheet (KCD) acts as the primary layer over which PG nanofibers were electrospun to act as the secondary layer. It is shown that this engineered bilayered nanofibrous scaffold material (KC-PG) should fulfill the functions of the extracellular matrix (ECM) by elucidating its function in vitro and in vivo. The bilayered nanofibrous scaffold was designed to exhibit improved physiochemical, biological and mechanical properties, with better swelling and porosity for enhanced oxygen permeability, and it also exhibits an acceptable antibacterial property to prevent infection at the wound site. The bilayered nanofibrous scaffold assists in better biocompatibility towards fibroblast and keratinocyte cell lines. The morphology of the nanofibrous scaffold aids increased cell adhesion and proliferation with cell material interactions. This was elucidated with the help of in vitro fluorescence staining against both cell lines. The bilayered KCD-PG nanofibrous scaffold material gives accelerated wound healing efficiency during in vivo wound healing. The results showed the regulation of growth factors with enhanced collagen synthesis, thereby helping in faster wound healing.

Synthesis, characterization and biological evaluation of chromen and pyrano chromen-5-one derivatives impregnated into a novel collagen based scaffold for tissue engineering applications

In this article, we describe the synthesis and biological evaluation of a novel 2-(methylamino)-3-nitro-4-(4-oxo-4H-chromen-3-yl) pyrano[3,2-c]chromen-5(4H)-one (CCN). It is also determined that CCN impregnated into the collagen scaffold has the potential to mimic the function of the extracellular matrix as a biomaterial in the field of tissue engineering. The series of pyrano[3,2-c]chromen-5(4H)-one derivatives (4a–4j), was analyzed by 1H NMR, 13C NMR, mass spectra and FTIR analysis. Compound 4c was confirmed by single crystal XRD studies. All the compounds were screened for antimicrobial activity against Gram positive, Gram negative bacteria and yeast. Among all the compounds, compound (4aCCN) showed activity against Gram positive and Gram negative bacteria, when compared to the synthesized compounds. Further, compound CCN was evaluated for cytotoxicity against MCF-7, Hep-2 and Vero cancer cell lines with IC50 values of 5.4 μg ml−1, 5.3 μg ml−1 and 68.4 μg ml−1 respectively. In addition, the results of flow cytometry and docking (PDBID: 1A27 with the ligand) studies supported the activity of the synthesized compound (4a). FTIR and NMR analysis of the CCN impregnated collagen scaffold were done to reveal the existence of the CCN molecule in the scaffold. The inherent property of the collagen scaffold was not significantly affected by the structure of the CCN molecule. The thermal and mechanical properties of the collagen scaffold impregnated with CCN molecules gives stability as well as supports the swelling. However, the COL-CCN scaffold showed an enhanced cell attachment and proliferation of NIH 3T3 fibroblast cells. Based on the results, the novel CCN molecule impregnated within a collagen scaffold has potential application as a biomaterial in tissue engineering.

Synthesis of highly interconnected 3D scaffold from Arothron stellatus skin collagen for tissue engineering application.

The substrate which is avidly used for tissue engineering applications should have good mechanical and biocompatible properties, and all these parameters are often considered as essential for dermal reformation. Highly interconnected three dimensional (3D) wound dressing material with enhanced structural integrity was synthesized from Arothron stellatus fish skin (AsFS) collagen for tissue engineering applications. The synthesized 3D collagen sponge (COL-SPG) was further characterized by different physicochemical methods. The scanning electron microscopy analysis of the material demonstrated that well interconnected pores with homogeneous microstructure on the surface aids higher swelling index and that the material also possessed good mechanical properties with a Youngs modulus of 0.89±0.2 MPa. Biocompatibility of the 3D COL-SPG showed 92% growth for both NIH 3T3 fibroblasts and keratinocytes. Overall, the study revealed that synthesized 3D COL-SPG from fish skin will act as a promising wound dressing in skin tissue engineering.

Biomimetic interconnected porous keratin-fibrin-gelatin 3D sponge for tissue engineering application.

The medicated wound dressing material with highly interconnected pores, mimicking the function of the extracellular matrix was fabricated for the promotion of cell growth. In this study, keratin (K), fibrin (F) and gelatin (G) composite scaffold (KFG-SPG) was fabricated by freeze drying technique and the mupirocin (D) drug was successfully incorporated with KFG-SPG (KFG-SPG-D) intended for tissue engineering applications. The fabrication of scaffold was performed without the use of any strong chemical solvents, and the solid sponge scaffold was obtained with well interconnected pores. The porous morphology of the scaffold was confirmed by SEM analysis and exhibited competent mechanical properties. KFG-SPG and KFG-SPG-D possess high level of biocompatibility, cell proliferation and cell adhesion of NIH 3T3 fibroblast and human keratinocytes (HaCaT) cell lines thereby indicating the scaffolds potential as a suitable medicated dressing for wound healing.

Evaluation of Stress-Induced Microbial Siderophore from Pseudomonas aeruginosa Strain S1 as a Potential Matrix Metalloproteinase Inhibitor in Wound Healing Applications

Abstract Matrix metalloproteinases (MMPs) are zinc-dependent proteolytic enzymes capable of causing various inflammatory and various degenerative diseases if over-expressed. The active site of these enzymes is a zinc binding motif which binds to the specific site on the substrate and induce degradation. Hence an inhibitor is required to form a complex with zinc motif which hampers the binding ability of MMPs. To obtain novel MMPs inhibitor for wound healing, the chelating activity of siderophore from the microbial source was focused. During screening for siderophore production, strain S1 produced the highest amount of siderophore in the minimal salts medium. The isolate was confirmed as Pseudomonas aeruginosa strain S1 based on 16S rRNA gene sequencing and phylogenetic analysis. The activity of the siderophore was assayed using chrome azurol sulphonate and purified by the chromatographic techniques. The structural evidence through Fourier transform infrared and nuclear magnetic resonance spectra revealed that the isolated siderophore is a catecholate type with the distinctive characters. The positive results of calcein and fluozin-3 assays indicate that siderophore could bind to divalent metal ions, namely Fe2+ and Zn2+. As the siderophore compound focused on wound healing property, the in vitro studies revealed the viability of NH3T3 fibroblast cells and its efficiency in matrix modulating was confirmed through gelatin zymogram.

Nanofibrous matrixes with biologically active hydroxybenzophenazine pyrazolone compound for cancer theranostics

The nanomaterial with the novel biologically active compounds has been actively investigated for application in cancer research. Substantial use of nanofibrous scaffold for cancer research with potentially bioactive compounds through electrospinning has not been fully explored. Here, we describe the series of fabrication of nanofibrous scaffold loaded with novel potential biologically active hydroxybenzo[a]phenazine pyrazol-5(4H)-one derivatives were designed, synthesized by a simple one-pot, two step four component condensation based on Michael type addition reaction of lawsone, benzene-1,2-diamine, aromatic aldehydes and 3-methyl-1-phenyl-1H-pyrazol-5(4H)-one as the substrates. The heterogeneous solid state catalyst (Fe (III) Y-Zeolite) could effectively catalyze the reaction to obtain the product with high yield and short reaction time. The synthesized compounds (5a-5p) were analyzed by NMR, FTIR and HRMS analysis. Compound 5c was confirmed by single crystal XRD studies. All the compounds were biologically evaluated for their potential inhibitory effect on anticancer (MCF-7, Hep-2) and microbial (MRSA, MTCC 201 and FRCA) activities. Among the compounds 5i exhibited the highest levels of inhibitory activity against both MCF-7, Hep-2 cell lines. Furthermore, the compound 5i (BPP) was evaluated for DNA fragmentation, flow cytometry studies and cytotoxicity against MCF-7, Hep-2 and NIH 3T3 fibroblast cell lines. In addition, molecular docking (PDB ID: 1T46) studies were performed to predict the binding affinity of ligand with receptor. Moreover, the synthesized BPP compound was loaded in to the PHB-PCL nanofibrous scaffold to check the cytotoxicity against the MCF-7, Hep-2 and NIH 3T3 fibroblast cell lines. The in vitro apoptotic potential of the PHB-PCL-BPP nanofibrous scaffold was assessed against MCF-7, Hep-2 cancerous cells and fibroblast cells at 12, 24 and 48h respectively. The nanofibrous scaffold with BPP can induce apoptosis and also suppress the proliferation of cancerous cells. We anticipate that our results can provide better potential research in nanomaterial based cancer research.

Fabrication of Arothron stellatus skin collagen film incorporated with Coccinia grandis as a durable wound construct

ABSTRACT The wound dressing material has been fabricated with the novel Arothron stellatus fish skin collagen scaffold loaded with a bioactive extract obtained from Coccinia grandis (CPE) and drug Ciprofloxacin (D). The collagen scaffold was fabricated from the nonedible solid waste of marine origin (a cheaper source of raw material). The in vitro fluorescence staining of cells and in vivo evaluation of the fabricated scaffold exhibited enhanced cell adhesion and proliferation and indicated a more efficient wound healing paradigm than that of the control and other treated groups, respectively. The increased collagen synthesis and re-epithelialization of wound are proved to be potential wound constructs. GRAPHICAL ABSTRACT

Dual-layered 3D nanofibrous matrix incorporated with dual drugs and their synergetic effect on accelerating wound healing through growth factor regulation

The by-product of the slaughter house was utilized for the development of promising regenerative wound dressing material. Currently, dual-layered nanofibrous spongy scaffold was fabricated for tissue engineering applications. Herein, Keratin (K)-Fibrin (F)-Gelatin (G) 3D sponge loaded with Mupirocin (M) was fabricated with the naturally derived materials from bovine origin using freeze drying method. Moreover, poly(3-hydroxybutyric acid) (P) and Gelatin(G) solution loaded with Curcumin (C) were electrospun to get the dual drug loaded dual-layered nanofibrous spongy 3D scaffold (KFG:M-PG:C). The fabricated biomaterial was assisted with physical, biological and mechanical property. The in vitro cell viability and fluorescence staining of NIH 3T3 and HaCaT cells assist in cell adhesion and proliferation of the dual-layered scaffold. Moreover, in vivo assessment using silicone splint animal model was employed. The nanofibrous surface aids in the migration of fibroblast for the increased the collagen deposition and granulation tissue formation. Nonetheless, the 3D spongy surface promotes the gaseous exchange and absorption of exudates. The fabricated KFG:M-PG:C scaffold has the ability to produce perusable material that can integrate with the host tissue. Overall, the three dimensional (3D) dual-layered nanofibrous spongy scaffold with synergistic effect of dual drugs prevents from infection and facilitates as highly durable substrate in tissue engineering application.