Browsing by Author "Ramakrishnan, R"

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    . Exploring the Potential of Alginate-Gelatin-Diethylaminoethyl Cellulose Fibrinogen based Bioink for 3D Bioprinting of Skin
    (Carbohydrate Polymer Technologies and Applications, 2022-06) Ramakrishnan, R; Kasoju, N; Raju, R; Geevarghese, R; Gauthaman, A; Bhatt, A
    Designing printable bioinks for 3D bioprinting capable of supporting cellular viability with post-printing functionality remains challenging. Native ECM offers several physical, chemical, and biological cues that are difficult to restore using only a single component. Herein, we have optimized a multicomponent-based bioink formulation comprising alginate (ALG), gelatin (GEL), diethylaminoethyl cellulose (DCEL) and fibrinogen (FIB), termed as ALG-GEL-DCEL-FIB bioink for potential application in bioprinting and biofabrication of skin tissue equivalents. The designed formulation was extensively studied for its printability, physico-chemical, rheological, and biocompatibility properties. Excellent printability, shape fidelity and cell-laden tissue equivalent printing were established using the RegenHu 3D Discovery Bioprinter. The human primary fibroblast and keratinocyte-laden bioprinted constructs exhibited good cell viability. Long term culture of 4 weeks comprising 5 days of air-liquid-interphase followed by 21 days of submerged culture produced biomimetic tissue histology in the ALG-GEL-DCEL-FIB bioink printed constructs. Specific epidermal-dermal marker expressions proving functionality were evident in immunohistochemical, biochemical and gene expression analysis. The ALG-GEL-DCEL-FIB bioink may be explored further for potential biofabrication and therapeutic applications.
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    Extracellular matrix-based combination scaffold for guided regeneration of large-area full-thickness rabbit burn wounds upon a single application
    (Journal of Biomedical Material Research Part B Applied Biomaterials, 2021-11) Ramakrishnan, R; Harikrishnan, VS; Anil, A; Sabareeswaran, A; Krishnan, LK
    Regeneration of large acute and chronic wounds is a concern worldwide. The present study evaluates wound healing competence of a completely human-origin, extracellular matrix (ECM)-based skin substitute/graft. It comprises cell-less amniotic membrane (AM), clinical-grade fibrin (FIB), and hyaluronic acid (HA) termed as AMFIBHA. The use of large-area third-degree rabbit burn wounds evaluated the product efficiency. The AMFIBHA induces hemostasis and permits suture-less positioning on the wound bed. In wet wounds, the AMFIBHA degrades and release biologically active molecules and guide cell migration, proliferation, and regeneration. The study demonstrated the effectiveness of this wound care product in terms of epithelial-dermal regeneration with angiogenesis. The study assessed injury-associated inflammation and different wound healing markers after 28 days of experiment and compared with both positive and negative controls-treated wounds. The regeneration of mature epidermis and dermis with rete pegs and hair follicle-like structure was evident upon a single application. The active involvement of host cells resulted in supple tissue formation. The ECM organization of AMFIBHA-treated tissue resulted in re-gain of mechanical properties comparable to native skin after 56 days. These guided regenerative outcomes reveal a promising translational value of the novel AMFIBHA skin substitute as an off-the-shelf product for clinical use.
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    Potential Skin Substitute of Biomimetic Proteins and Terpolymer with Proven Immuno-Compatible and Biodegradable Properties
    (Paripex - Indian Journal of Research, 2020-02) Ramakrishnan, R; Mohanan, PV; Krishnan, LK
    Non–immunogenic matrix with desirable biological and mechanical properties could be a valuable graft to treat large area acute wounds and non–healing chronic wounds. In this study, an electrospun biodegradable polymer scaffold composed of a terpolymer PLGC [poly(lactide–glycolide–caprolactone)] incorporated with a fiin–hyaluronic acid (HA) based biomimetic composite (PLGCFIBHA) was evaluated. An appropriate mechanical properties, fioblast growth potential, biodegradability and absence of immune response were established. A significantly increased fioblast attachment and proliferation was observed with optimal physical properties. Intracutaneous (intradermal) reactivity test and guinea pig maximization test was negative as per requirements of ISO–10993–10:2010(E): biological evaluation of medical devices part 10 proving immunocompatible nature. In vitro and in vivo studies showed suitable degradability for skin tissue engineering purposes. Hence the faicated scaffold PLGCFIBHA is advocated as a potential candidate for the engineering of dermal tissue which can be used as an off–the–shelf product.
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    Reinforcement of amniotic membrane with fibrin coated poly- [Lactide-co-Glycolide-co-Caprolactone] terpolymer containing silver nanoparticles for potential wound healing applications
    (International Journal of Polymeric Materials and Polymeric Biomaterials, 2019-06) Ramakrishnan, R; Krishnan, LK; Nair, RP; Krishnan, KV
    Human amniotic membrane (AM) is reported to possess remarkable therapeutic potential for wound healing due to its inherent bioactive and biocompatible properties. However, poor handling characteristics have limited its clinical use on wounds to a great extent. Reinforcement of AM with a biodegradable polymer is therefore expected to provide adequate mechanical and handling nature. Poly-[Lactide-co-Glycolide-co-Caprolactone] terpolymer incorporated with silver nanoparticles and coated with fibrin is used in this study to reinforce the membrane which delivers bioactive molecules to the wound site without wound contraction. This combination scaffold exhibited desirable properties with excellent biocompatibility for potential use in wound healing applications.