Browsing by Author "G. Taru Sharma"

Now showing 1 - 7 of 7

Biomaterials and Scaffolds in Stem Cell Therapy
(Springer Nature, 2022) Mukesh Kumar Bharti; Vikash Chandra; G. Taru Sharma
In the recent past, stem cell therapy has considerably revolutionized regenerative therapy; however, still, it is not perfect for treating diseases due to several limitations like transplantation of stem cells alone exhibits low therapeutic efficacy due to poor viability and regenerative activity of transplanted cells. There is a high scope for the improvement of ex vivo stem cell culture and its delivery system. Growth factors and cytokines regulate stem cell proliferation and differentiation; besides this, they also require biophysical cues at their niche. To overcome these limitations, techniques of tissue engineering use scaffolds. Scaffold opens new avenues for producing engineered tissue substitutes and thus by quality organ repair. Biophysical signals from bioscaffolds such as mechanical forces, nanotopography, stiffness of the matrix, and surface features of the biomaterial influence stem cells’ fate. Several types of scaffolds are being used derived from natural biomaterials or synthetic materials having their own merits and demerits. Biodegradability and biologically active properties are the major advantages of natural bioscaffolds over synthetic scaffolds. However, the major drawback of natural scaffolds is the risk of carrying cross-contaminated from the sources. Technologies evolved to mold the biomaterials into three-dimensional (3D) scaffolds to simulate tissue architecture to promote cell proliferation and differentiation. Combining stem cell technologies with biomaterial-based scaffolds enhance stem cell viability, differentiation, and therapeutic efficacy. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2021.
Comparative study on characterization and wound healing potential of goat (Capra hircus) mesenchymal stem cells derived from fetal origin amniotic fluid and adult bone marrow
(Elsevier B.V., 2017) M.D. Pratheesh; Pawan K. Dubey; Nitin E. Gade; Amar Nath; T.B. Sivanarayanan; D.N. Madhu; Anjali Somal; Indu Baiju; T.R. Sreekumar; V.L. Gleeja; Irfan A. Bhatt; Vikash Chandra; Amarpal; Bhaskar Sharma; G. Saikumar; G. Taru Sharma
Caprine amniotic fluid (cAF) and bone marrow cells (cBM) were isolated, expanded and phenotypically characterized by mesenchymal stem cells (MSCs) specific cell surface markers. Both cell types were compared for multilineage differentiation potential by flow cytometry using specific antibodies against lineage specific markers. Furthermore, in vitro expanded cAF-MSCs showed higher expression of trophic factors viz. VEGF and TGF-β1 as compared to cBM-MSCs. Full-skin thickness excisional wounds created on either side of the dorsal midline (thoracolumbar) of New Zealand White rabbits were randomly assigned to subcutaneous injection of either fetal origin cAF-MSCs (n = 4) or adult cBM-MSCs (n = 4) or sterile PBS (control, n = 4). The rate of wound closure was found faster (p < 0.05) in cAF-MSCs treated wounds as compared with cBM-MSCs and PBS treated wounds especially on 21st day post-skin excision. Histomorphological examination of the healing tissue showed that wound healing was improved (p < 0.05) by greater epithelialization, neovascularization and collagen development in cAF-MSCs as compared to cBM-MSCs and PBS treated wounds. © 2017 Elsevier Ltd
Evaluation of canine bone marrow-derived mesenchymal stem cells for experimental full-thickness cutaneous wounds in a diabetic rat model
(Taylor and Francis Ltd., 2021) Deepika Bist; A.M. Pawde; Amarpal; Prakash Kinjavdekar; Reena Mukherjee; K.P. Singh; Med Ram Verma; Khan Sharun; Amit Kumar; Pawan K. Dubey; Divya Mohan; Amit Verma; G. Taru Sharma
Background: The wound healing potential of canine bone marrow-derived mesenchymal stem cells (BMSCs) was evaluated in the excisional wound of streptozotocin-induced diabetic rats. Research design and methods: Xenogenic BMSCs were collected aseptically from the iliac crest of healthy canine donors under general anesthesia. Full-thickness experimental wounds (20 × 20 mm2) on the dorsum of forty-eight adult healthy Wistar white rats. The wounds were assigned randomly to three treatment groups: PBS (Group A) or BMSCs (Group B) injected into the wound margins on days 0, 7, and 14 or BMSCs (Group C) injected into the wound margins on days 7, 14, and 21 post-wounding. The degree of wound healing was evaluated based on macroscopical, hemato-biochemical, histopathological, and histochemical parameters. Results: The results indicated granulation tissue formation with reduced exudation and peripheral swelling in the treatment groups compared to the control group A. Similarly, the degree of wound contraction was significantly higher in groups B and C animals than group A on days 14 and 21 post-wounding. The transplantation of BMSCs resulted in early drying of wounds, granulation tissue appearance, and enhanced cosmetic appearance. Conclusion: The histopathological, histochemical, and gross findings suggested the therapeutic potential of xenogeneic mesenchymal stem cell therapy in managing diabetic wounds. Abbreviations: BMSCs-bone marrow-derived mesenchymal stem cells, PBS-phosphate-buffered saline, MSCs-mesenchymal stem cells, FBS-fetal bovine serum, ECM-extracellular matrix. © 2021 Informa UK Limited, trading as Taylor & Francis Group.
Evaluation of persistence and distribution of intra-dermally administered PKH26 labelled goat bone marrow derived mesenchymal stem cells in cutaneous wound healing model
(Springer Netherlands, 2017) M.D. Pratheesh; Nitin E. Gade; Amar Nath; Pawan K. Dubey; T.B. Sivanarayanan; D.N. Madhu; T.R. Sreekumar; Amarpal; G. Saikumar; G. Taru Sharma
The current study was designed to study the persistence and distribution of caprine bone marrow derived mesenchymal stem cells (cBM-MSCs) when administered intra-dermally in experimentally induced cutaneous wounds in rabbits. MSC’s from goat bone marrow were isolated and their differentiation potential towards adipogenic and osteogenic lineages were assayed in vitro. The isolated cells were phenotypically analysed using flow cytometry for the expression of MSC specific matrix receptors (CD73, CD105 and Stro-1) and absence of hematopoietic lineage markers. Further, these in vitro expanded MSCs were stained with PKH26 lipophilic cell membrane red fluorescent dye and prepared for transplantation into cutaneous wounds created on rabbits. Five, 2 cm linear full thickness skin incisions were created on either side of dorsal midline of New Zealand white rabbits (n = 4). Four wounds in each animal were implanted intra-dermally with PKH26 labelled cBM-MSCs suspended in 500 µl of Phosphate Buffer Saline (PBS). Fifth wound was injected with PBS alone and treated as negative control. The skin samples were collected from respective wounds on 3, 7, 10 and 14 days after the wound creation, and cryosections of 6 µM were made from it. Fluorescent microscopy of these cryosections showed that the PKH26 labelled transplanted cells and their daughter cells demonstrated a diffuse pattern of distribution initially and were later concentrated towards the wound edges and finally appeared to be engrafted with the newly developed skin tissues. The labelled cells were found retained in the wound bed throughout the period of 14 days of experimental study with a gradual decline in their intensity of red fluorescence probably due to the dye dilution as a result of multiple cell division. The retention of transplanted MSCs within the wound bed even after the complete wound healing suggests that in addition to their paracrine actions as already been reported, they may have direct involvement in various stages of intricate wound healing process which needs to be explored further. © 2017, Springer Science+Business Media Dordrecht.
Exploration of immunomodulatory mechanism of caprine Wharton's jelly derived mesenchymal stem cells
(Academic Press Inc., 2024) Indu Baiju; Mukesh Kumar Bharti; Anjali Somal; Sriti Pandey; Irfan A. Bhat; Anand Joseph; Vikash Chandra; G. Taru Sharma
The present study was aimed to explore the possible mechanisms by which caprine Wharton's jelly-derived MSCs (WJ-MSCs) perform their immunomodulatory function. WJ-MSCs were isolated through explants culture and characterized as per ISCT criteria using culture behavior, expression of surface markers by PCR, FACS and immunocytochemical localization (ICC), trilineage differentiation potential etc. Secretory behavior for important biomolecules (IDO, TGFβ1, VEGF, IL6) was evaluated by ICC and western blot assay. Cell-to-cell communication was studied by culturing cells in cell–cell contact and trans-well system. The MSCs when co-cultured with activated Tc and Th cells, down-regulation of T cell cytokine as well as upregulation of immunomodulatory factors (VEGF A, IL10, IL6, IDO, iNOS, PTGS2, HGF, TGFβ, CXCL10, CXCL11) was noticed in both cell–cell contact and trans-well culture system which was significantly higher in cell–cell contact system. Trilineage differentiation of MSCs showed significant upregulation of MHC I (CAHI) and MHC II (CLA DRB3) molecules suggesting better clinical applications of MSCs without differentiation to avoid immune rejection. It can be concluded that WJ-MSCs perform their immunomodulation through the secretion of a battery of biomolecules and work in both cell–cell contact manner and through their secretome. © 2024 Elsevier Inc.
Expression profile of adhesion molecules in blastocyst vis-a-vis uterine epithelial cells
(Elsevier Inc., 2021) Sriti Pandey; H. Lakshmi Devi; Irfan Ahmad Bhat; B. Indu; Mukesh Kumar Bharti; Uffaq Shabir; Bilal Ahmad Peer; Vikash Chandra; G. Taru Sharma
Models using in vitro produced buffalo embryos and in vitro cultured uterine epithelial cells (UECs) may be useful in understanding the intricacies of embryo-uterine cross talk. In the present study, buffalo UECs were obtained from slaughterhouse derived non-gravid uterus. UECs monolayer was treated with steroids (10pg/ml estradiol for 24h and 3.14 ng/ml progesterone for another 5 days). In vitro produced buffalo blastocysts were co-cultured over steroid treated UECs monolayer and at 72 h of co-culture, embryo attachment rate was higher in UECs treated with steroids (71.86% vs. 26.55%) while no attachment was observed on plastic surface. Naturally hatched or assisted hatched blastocysts were co-cultured over UECs monolayer treated with 3.14ng/ml progesterone (P4), or without any treatment for 72 h and the effect of co-culture on the expression profile of adhesion related biomolecules was analyed in UECs and blastocysts. Cultured UECs and blastocysts cultured in embryo culture media were considered as control. It was observed that the expression of MUC1 in UECs was significantly (p < 0.05) higher in control group than treatment groups. The relative mRNA abundance of integrins and osteopontin was significantly (p < 0.05) higher in UECs and blastocysts of treatment groups than control group. Expression of IFN-τ was significantly higher (p < 0.05) in embryos co-cultured with UECs than other treatment groups. It can be concluded that P4 supplementation is required for the modulation of adhesion molecules and co-culture of blastocysts and UECs together affect the expression of adhesion molecules both in blastocyts and in UECs. © 2021 Elsevier Inc.
Isolation and propagation of classical swine fever virus in porcine Wharton’s Jelly mesenchymal stem cells
(Taylor and Francis Ltd., 2022) Neelam R. Tomar; Irfan A. Bhat; Mukesh K. Bharti; Jeny K. John; Veena Sharma; Vikash Chandra; G. Taru Sharma; G. Saikumar
Classical Swine Fever (CSF) is an extremely infectious and deadly disease of pigs and wild boars caused by the CSF virus (CSFV) which is a member of the Pestivirus genus and the family Flaviviridae. This study was designed to detect the permissibility and replication of CSFV in mesenchymal stem cells (MSCs) monolayer derived from Porcine Wharton’s jelly. Porcine Wharton’s jelly MSCs (pWJ-MSCs) were ex vivo expanded and propagated for more than 81 generations and third passage pWJ-MSCs were characterized as per standard criteria i.e., growth characteristics, trilineage differentiation potential and molecular characterization for pluripotency and stem cell surface markers. Porcine WJ tissue samples found negative for CSFV by RT-PCR test were processed further for the isolation of pWJ-MSCs and CSFV was propagated over the characterized pWJ-MSCs monolayer. No cytopathic effect was observed, which was consistent with non-cytopathic nature of CSFV. The replication of CSFV in pWJ-MSCs was affirmed by RT-PCR and demonstration of viral antigen in the cytoplasm of virus infected cells by immuno-staining technique. In total, three different CSFV isolates were propagated in pWJ-MSCs. Primary pWJ-MSCs permitted CSFV replication to good titer. To the best of our information, this is the first ever report of isolation of CSFV in pWJ-MSCs. © 2020 Taylor & Francis Group, LLC.