Browsing by Author "Krishnan, L"

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    Design of a Biomimetic Niche for Adult Progenitor Cell Selection and Differentiation
    (ADULT STEM CELL STANDARDIZATION, 2011) Krishnan, L
    Human peripheral blood mononuclear cell (PBMNC) contains a mixture of progenitor cells with potential to differentiate in to a wide range of lineages. Therefore, use of autologous progenitors from blood circulation could be a good treatment option, however; due to the limited numbers available, in vitro cell expansion may be crucial. Isolation of lineage committed progenitors for regenerative medicine is required because other contaminating cells may have adverse effect on the tissue of interest, however; obtaining pure population is often difficult due to the absence of appropriate surface markers. Therefore, anchorage dependent circulating progenitors could be allowed to home to cell-specific biomimetic niche on which only the specific cell type may home, survive and differentiate to the desired cell and multiply to get them in larger numbers. In the proof-of-concept presented here, each culture was started from peripheral blood mononuclear cells (PBMNC) on cell specific matrix compositions which consisted of adhesive proteins, growth factors and glycosaminoglcans. Three cell types of interest were endothelial progenitor cells (EPCs), smooth muscle cell progenitors (SMPCs) and neural progenitor cells (NPC). Differentiation of EPC into endothelial cells (EC), SMPC to smooth muscle cells (SMCs) and NPC to neurons were evaluated by their respective morphological characteristics and cell specific immuno cytochemical signatures. The EC that differentiated from EPC expressed vWF & endocytosed AcLDL, SMC that differentiated from SMPC expressed SMA and calponin. The neurons that were developed from NPCs expressed an early marker (beta-tubulin III) and a terminal marker (MAP-2). This chapter puts forward an approach of employing cell-specific niche for differentiation of PBMNC fraction of human blood into EC, SMC and neurons. The usefulness of the concept is established to harvest and multiply these progenitors in vitro to obtain sufficient number of required cell types which may find application in regenerative medicine.
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    Evaluation of Albuminated Curcumin as Soluble Drug Form to Control Growth of Cancer Cells in Vitro
    (Journal of Cancer Therapy., 2015-04) Thomas, C; Pillai, LS; Krishnan, L
    Curcumin (Curc) is well known for its anticancer activity, but its poor solubility in aqueous medium is a major concern for little therapeutic outcome. Therefore, the effort to improve its bioavailability is a major research interest. The current study aimed at conjugation of Curc to serum albumin (Alb) to increase aqueous solubility of the former without affecting its drug action on cancer cell lines and primary cells in culture. Conditions for preparation of albumin-curcumin (Alb-Curc) conjugate were standardized to obtain pure and stable drug. The product was obtained in sufficient quantity to test its effect on cells in culture at different doses. Briefly, the conjugate was prepared by mixing Curc dissolved in DMSO with the Alb dissolved in phosphate buffered saline; conjugate was purified by gel filtration chromatography and was analyzed using UV-Vis spectroscopy for characteristic peaks of both molecules. The conjugate was added to culture medium to identify the effect of conjugate on cell cycling and apoptosis. Albuminated curcumin that showed 100-fold higher solubility than free Curc was stable and inhibitory to proliferation, induced cell cycle arrest and apoptosis. The conjugate showed apoptotic effects on endothelial cells indicating its anti angiogenic property. Primary fibroblast growth was also inhibited but at the higher dose. The in vitro results suggest that Alb-Curc which is free of insoluble native drug may find application in cancer therapy after appropriate in vivo evaluations.
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    Feasibility of Dermal Substitute Construction on Hybrid Scaffold Made of Poly(epsilon-caprolactone) and Bio Mimetic Fibrin Composite
    (JOURNAL OF BIOMATERIALS AND TISSUE ENGINEERING, 2014) Nair, RP; Krishnan, K; Krishnan, L
    Non-healing wounds can be a major problem in diabetic and burn victims. Identified causes of chronic wound formation include angiopathy, neuropathy, infection and loss of extra cellular matrix (ECM) due to increased protease action. Currently available conventional therapy is not efficient enough to promote wound healing. Tissue-engineered skin substitutes are now considered a better strategy for chronic wound management. Non-toxic scaffolds on which cells grow and replace the lost ECM are important components for tissue engineering. Degradable polymers are preferred because they may be absorbed by the time the tissue regenerate. We attempted to fabricate a degradable membrane-like and porous poly(epsilon-caprolactone)(PCL) scaffold to favour in-growth of cells, penetration of nutrients and oxygen. Modifying porous PCL membrane using a fibrin-based bionnimetic matrix could promote cell attachment and growth of tissue. The benefit of such a hybrid scaffold for long-term cell growth and ECM deposition, parallel to the degradation of polymer and diminution of mechanical property, was evaluated. We concluded that the hybrid scaffold we developed is suitable for dermal tissue construction in vitro. When grown on the hybrid scaffold, cells synthesized and deposited insoluble ECM proteins, thus proposing better epithelialization and angiogenesis when such dermal tissue constructs are implanted for treating non-healing chronic wounds.