The Yin and Yang of Immunity in Stem Cell Decision Guidance in Tissue Ecologies: An Infection Independent Perspective.

The impact of immune system and inflammation on organ homeostasis and tissue stem cell niches in the absence of pathogen invasion has long remained a conundrum in the field of regenerative medicine. The paradoxical role of immune components in promoting tissue injury as well as resolving tissue damage has complicated therapeutic targeting of inflammation as a means to attain tissue homeostasis in degenerative disease contexts. This confound could be resolved by an integrated intricate assessment of cross-talk between inflammatory components and micro- and macro-environmental factors existing in tissues during health and disease. Prudent fate choice decisions of stem cells and their differentiated progeny are key to maintain tissue integrity and function. Stem cells have to exercise this fate choice in consultation with other tissue components. With this respect tissue immune components, danger/damage sensing molecules driving sterile inflammatory signaling cascades and barrier cells having immune-surveillance functions play pivotal roles in supervising stem cell decisions in their niches. Stem cells learn from their previous damage encounters, either endogenous or exogenous, or adapt to persistent micro-environmental changes to orchestrate their decisions. Thus understanding the communication networks between stem cells and immune system components is essential to comprehend stem cell decisions in endogenous tissue niches. Further the systemic interactions between tissue niches integrated through immune networks serve as patrolling systems to establish communication links and orchestrate micro-immune ecologies to better organismal response to injury and promote regeneration. Understanding these communication links is key to devise immune-centric regenerative therapies. Thus the present review is an integrated attempt to provide a unified purview of how inflammation and immune cells provide guidance to stem cells for tissue sculpting during development, organismal aging and tissue crisis based on the current knowledge in the field.

Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions.

Bone marrow-derived mesenchymal stromal cells (BM-MSCs) are gaining increasing importance in the field of regenerative medicine. Although therapeutic value of MSCs is now being established through many clinical trials, issues have been raised regarding their expansion as per regulatory guidelines. Fetal bovine serum usage in cell therapy poses difficulties due to its less-defined, highly variable composition and safety issues. Hence, there is a need for transition from serum-based to serum-free media (SFM). Since SFM are cell type-specific, a precise analysis of the properties of MSCs cultured in SFM is required to determine the most suitable one. Six different commercially available low serum/SFM with two different seeding densities were evaluated to explore their ability to support the growth and expansion of BM-MSCs and assess the characteristics of BM-MSCs cultured in these media. Except for one of the SFM, all other media tested supported the growth of BM-MSCs at a low seeding density. No significant differences were observed in the expression of MSC specific markers among the various media tested. In contrary, the population doubling time, cell yield, potency, colony-forming ability, differentiation potential, and immunosuppressive properties of MSCs varied with one another. We show that SFM tested supports the growth and expansion of BM-MSCs even at low seeding density and may serve as possible replacement for animal-derived serum.

Evaluation of vascular endothelial growth factor - A release from platelet-rich fibrin, platelet-rich fibrin matrix, and dental pulp at different time intervals.

BACKGROUND: Lack of collateral blood supply and the low compliance environment of pulp makes its healing, a challenge. Vascular endothelial growth factor (VEGF) is a primary angiogenic growth factor. Knowledge of dose and time-dependent expression of VEGF from platelet concentrates, namely platelet-rich fibrin (PRF) and PRF matrix (PRFM), along with vital pulp can aid in developing strategies to improve the outcome of vital pulp therapy and regenerative procedures. Hence, the aim of this study was to compare VEGF kinetics of PRF, PRFM, and dental pulp. MATERIALS AND METHODS: The PRF, PRFM, and vital dental pulp were placed in culture media for a week; the supernatant was collected from these samples at days 1, 4, and 7. VEGF-A expression was evaluated using ELISA and compared with the weight of the sample so as to quantify the release of VEGF-A per milligram of sample. RESULTS: PRF exhibited maximum VEGF-A release on day 4 and was sustained till day 7. In contrast, PRFM and dental pulp showed no significant release of VEGF-A till day 7. However, on day 7, there was a rapid increase in VEGF-A expression from dental pulp that was comparable to PRF. On comparing the release of VEGF-A per milligram of tissue, pulp exhibited the maximum values. CONCLUSION: Among the platelet concentrates, differential expression of VEGF-A was superior in PRF. The use of PRF in partial pulpitis should be explored in order to restore pulp vascularity and hasten pulpal healing.

Evaluation of Histological and pH Changes in Platelet-Rich Fibrin and Platelet-Rich Fibrin Matrix: A In vitro Study.

BACKGROUND: The autologous platelet concentrates (PCs), such as platelet-rich fibrin (PRF) and platelet-rich fibrin matrix (PRFM), are processed through different centrifugation protocols, which can affect their biological properties and in turn influence treatment outcome. The pH value can influence the process of wound healing directly or indirectly. Hence, a study was conducted to evaluate fibrin network pattern, initial pH of obtained matrix, and its changes during degradation matrix and to determine platelet and leukocyte count in PRF and PRFM. MATERIALS AND METHODS: Blood from the volunteers was collected in blood vacutainers for processing PRF and PRFM. It was centrifuged as per the standard protocol. Serum from PRF and PRFM was subjected for analysis of platelet and leucocyte concentration using Hemo-Analyzer. The pH of PRF and PRFM were evaluated over 5 days using a pH meter. PRF and PRFM were subjected to histological and scanning electron microscopic analysis. RESULTS: There was no significant variation in the platelet and leukocyte count between PRF and PRFM. A steady rise in the level of pH with respect to PRFM was observed, whereas there was a decrease in the pH levels in PRF. Fibrin network was denser in PRF compared to PRFM. CONCLUSION: The PCs undergo variation in pH upon degradation. Formation of fibrin matrix is influenced by the method of preparation. Fibrin pattern is crucial to facilitate adhesion of cells and transport of nutrients to enable proliferation and differentiation of mesenchymal cells and better wound healing.

Regenerative therapy for hippocampal degenerative diseases: lessons from preclinical studies.

Increase in life expectancy has put neurodegenerative diseases on the rise. Amongst these, degenerative diseases involving hippocampus like Alzheimer's disease (AD) and temporal lobe epilepsy (TLE) are ranked higher as it is vulnerable to excitotoxicity induced neuronal dysfunction and death resulting in cognitive impairment. Modern medicines have not succeeded in halting the progression of these diseases rendering them incurable and often fatal. Under such scenario, regenerative studies employing stem cells or their by-products in animal models of AD and TLE have yielded encourageing results. This review focuses on the distinct cell types, such as hippocampal cell lines, neural precursor cells, embryonic stem cells derived neural precursor cells, induced pluripotent stem cells, induced neurons and mesenchymal stem cells, which can be employed to rescue hippocampal functions in neurodegenerative diseases like AD and TLE. Besides, the divergent mechanisms through which cell based therapy confer neuroprotection, current impediments and possible improvements in stem cell transplantation strategies are discussed. Authors are aware of the voluminous literature available on this issue and have made a sincere attempt to put forth the current status of research in the field of cell based therapy concurrently discussing the promise it holds for combating neurodegenerative diseases like AD and TLE in the near future. Copyright © 2015 John Wiley & Sons, Ltd.

Conditioned Medium Reconditions Hippocampal Neurons against Kainic Acid Induced Excitotoxicity: An In Vitro Study.

Stem cell therapy is gaining attention as a promising treatment option for neurodegenerative diseases. The functional efficacy of grafted cells is a matter of debate and the recent consensus is that the cellular and functional recoveries might be due to "by-stander" effects of grafted cells. In the present study, we investigated the neuroprotective effect of conditioned medium (CM) derived from human embryonic kidney (HEK) cells in a kainic acid (KA) induced hippocampal degeneration model system in in vitro condition. Hippocampal cell line was exposed to KA (200 µM) for 24 hrs (lesion group) whereas, in the treatment group, hippocampal cell line was exposed to KA in combination with HEK-CM (KA + HEK-CM). We observed that KA exposure to cells resulted in significant neuronal loss. Interestingly, HEK-CM cotreatment completely attenuated the excitotoxic effects of KA. In HEK-CM cotreatment group, the cell viability was ~85-95% as opposed to 47% in KA alone group. Further investigation demonstrated that treatment with HEK-CM stimulated the endogenous cell survival factors like brain derived neurotrophic factors (BDNF) and antiapoptotic factor Bcl-2, revealing the possible mechanism of neuroprotection. Our results suggest that HEK-CM protects hippocampal neurons against excitotoxicity by stimulating the host's endogenous cell survival mechanisms.