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Joshi L.,Bhagwan Mahavir Medical Research Center | Chelluri L.K.,Transplant Immunology and Stem Cell Laboratory | Gaddam S.,Bhagwan Mahavir Medical Research Center | Gaddam S.,Osmania University
Archivum Immunologiae et Therapiae Experimentalis | Year: 2015

Multi-drug-resistant (MDR) tuberculosis is a major public health problem worldwide. Drug resistance arises due to non-compliance of antibiotic therapy. Herein, we explored the therapeutic options available ranging from conservative treatment approaches to alternate adjunct therapies such as mesenchymal stromal cell (MSC) therapy interventions. It is attractive to understand the scientific rationale of using cells as drugs, in particular mesenchymal stem/stromal cells. The review dwells and attempts to analyze the mechanistic approaches of the current treatment modalities to modern therapies. MSCs have demonstrated profound capacity to regenerate and repair. They appear to modulate that the activities of dendritic cells regulate T cells, both in vivo and in vitro. While there seems to be some benefit of such therapies, its use warrants further research. The merits and de-merits of autologous therapy/allogeneic therapy are ill understood. The challenges of requirement of large number of cells for infusion, the route of administration, choice of timing are complex issues that need to be addressed. Furthermore, the host immune responses, environmental factors and epigenetic mechanisms compound the problem. Although, clinical studies are being performed using autologous MSCs in different inflammatory models, it is important that such an intervention should be based on sound scientific rationale. The current review examines the immunomodulatory properties of MSCs, its interactions with other cell types, in assessing the basis for autologous/allogeneic cell-based therapies in the treatment of XDR/MDR tuberculosis. © 2015, L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland. Source


Chelluri L.K.,Transplant Immunology and Stem Cell Laboratory | Prasad C.,A. P. Chest Hospital | Gokhale A.G.,Global Hospitals
Internet Journal of Infectious Diseases | Year: 2010

Tumor necrosis factor-alpha and interferon-gamma are two important pleiotropic inflammatory cytokines that perpetuate tissue damage after increased stress and in the presence of excess glucocorticoids with respect to tuberculosis (TB) infection. Activated macrophage dysfunction leads to granuloma formation, cavities, and fibrosis. Current treatment strategies are not amenable to long-term compliance, contraindications, increased relapse rates, and compounding drug resistance. A natural supplement devoid of the abovementioned complications needs to be developed and checked for healthy immune reconstitution targeting dendritic cell therapy. The preliminary prospective study in five TB subjects was assessed objectively using ELISA for TNF-alpha and interferon-gamma levels. The PBMCs were mixed and cultured for the effect of allogenic bone marrow-derived mesenchymal stem cell interactions and secretory cytokines in the culture supernatant. The TNF-alpha levels on day five with an MSC dose of 105 cells/ml for 1×106 cells/ml of PBMCs of diseased patients were significantly decreased while there was no change in IFN-gamma levels. The variable dose of MSC appears to have had a significant impact on the cytokine milieu in vitro. The results are encouraging in that there is possibility for effective immune modulation using mesenchymal stem cells in infection-mediated inflammation.Keywords: Mesenchymal therapy; Pulmonary TB infection; Cytokines; Immunomodulation; Mesenchymal stem cell dose. Source


Debnath T.,Transplant Immunology and Stem Cell Laboratory | Ghosh S.,Indian Institute of Chemical Technology | Potlapuvu U.S.,Transplant Immunology and Stem Cell Laboratory | Kona L.,Global Hospitals | And 4 more authors.
PLoS ONE | Year: 2015

Applied tissue engineering in regenerative medicine warrants our enhanced understanding of the biomaterials and its function. The aim of this study was to evaluate the proliferation and differentiation potential of human adipose-derived stem cells (hADSCs) grown on chitosan hydrogel. The stability of this hydrogel is pH-dependent and its swelling property is pivotal in providing a favorable matrix for cell growth. The study utilized an economical method of cross linking the chitosan with 0.5% glutaraldehyde. Following the isolation of hADSCs from omentum tissue, these cells were cultured and characterized on chitosan hydrogel. Subsequent assays that were performed included JC-1 staining for the mitochondrial integrity as a surrogate marker for viability, cell proliferation and growth kinetics by MTT assay, lineage specific differentiation under two-dimensional culture conditions. Confocal imaging, scanning electron microscopy (SEM), and flow cytometry were used to evaluate these assays. The study revealed that chitosan hydrogel promotes cell proliferation coupled with > 90% cell viability. Cytotoxicity assays demonstrated safety profile. Furthermore, glutaraldehyde cross linked chitosan showed < 5% cytotoxicity, thus serving as a scaffold and facilitating the expansion and differentiation of hADSCs across endoderm, ectoderm and mesoderm lineages. Additional functionalities can be added to this hydrogel, particularly those that regulate stem cell fate. © 2015 Debnath et al. Source


Debnath T.,Transplant Immunology and Stem Cell Laboratory | Shalini U.,Transplant Immunology and Stem Cell Laboratory | Kona L.K.,Global Hospitals | Vidya Sagar J.V.S.,Aware Global Hospitals | And 3 more authors.
Journal of Arthroscopy and Joint Surgery | Year: 2015

Aim: Alternate strategies to regenerate the damaged tissue require exogenous supply of several chondrocyte implants. There are inherent challenges to optimize an appropriate tissue culture methodology that can aid in enrichment of chondrocytes. The aim of the study was to explore the differentiation potential, expansion and growth kinetics of the human adipose derived stem cells (hADSCs) in alginate microspheres in comparison to chondrogenesis from the cartilage tissue. Methods: Isolated hADSCs and cartilage derived chondrocytes were cultured and characterized. The distribution of stem cells within alginate bead was imaged by scanning electron microscopy (SEM). Encapsulated hADSCs were monitored for their cell viability, cell proliferation and apoptosis via JC-1 staining, MTT assay and Annexin V assays respectively. The alginate cell constructs were analyzed for chondrogenic gene expression by RT-PCR. Results: The chondrocyte pellet culture from cartilage demonstrated lower growth potential as compared to alginate encapsulation. hADSCs were successfully encapsulated within alginate matrix with >80% cell viability. Apoptotic assays provided safety profile for the alginate during cell growth. The up-regulation of cartilage specific genes like TGF-β, collagen type-X, COMP was observed during the entire period of culture. Conclusion: The chondrogenesis in pellet culture from cartilage tissue conserved the chondrocyte phenotype better with rich GAG polysaccharides. However, owing to an enriched chondrocyte requirement, alginate as a scaffold design would aid in the treatment of large focal defects. © 2015 International Society for Knowledge for Surgeons on Arthroscopy and Arthroplasty. Source

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