Evaluation the ability of acellular ovine small intestine submucosa to load and release of mineral pitch and its anti-inflammatory effects

Introduction
Inflammation and subsequent events are complex biological processes that occur in response to injury or infection and involve various inflammatory mediators such as cytokines and chemokines (Orman et al. 2011). In conditions such as severe burns, the persistent inflammatory response is induced, leading to increased infection and sepsis, which in turn increased the risk of multi-organ failure and death (Finnerty et al. 2009). The burn injury dramatically increased macrophages exposed to lipopolysaccharide (LPS), increased tumor necrosis factor-a (TNF-a) and interleukin-1b (IL-1b) levels, and decreased IL-10 level.

(Li et al. 2016). In addition to these factors, local mediators such as reactive oxygen species (ROS) and nitric oxide are also secreted in burns (Ono et al. 1995; Summer et al. 2008; Evers et al. 2010). Despite progress in many aspects of burn healing over the past decades that have resulted in reduced mortality and morbidity, general inflammatory condition and secondary infection still remain serious problems of severe burns (Church et al. 2006; Ribeiro et al. 2010; Leclerc etal.2011).Theresultsofinvivostudies show that stem cells are both capable of reducing proinflammatory cytokine release and nitric oxide production and enhancing the regulation of anti-inflammatory cytokines (Weil et al. 2009). The extent of the immunosuppressive function of stem cells, is induced by activated immune cells and cytokines, is greater with human umbilical cord mesenchymal stem cells (hUC-MSCs) than with other MSCs (Prasanna et al. 2010). Recent studies showed that the release of soluble paracrine factors from hUC-MSCs can regulate the expression of inflammatory factors and activate the cells to secret kinds of bioactive factors, that regulated inflammatory and immune responses, and finally limit burn-induced inflammation (da Silva Meirelles et al. 2009; Li et al. 2016). Recently, the use of traditional herbal drugs for wound healing has attracted the attention of scientific circles, as about one-third of all traditional medicines are used to treat wounds and skin disorders (Mantle et al. 2001; Houghton et al. 2005; Annan and Houghton 2008). Mineral pitch (MP), also known as shilajit, salajit, shilajatu, mumie, or mummiyo is a herbo-mineral drug, which comes from layers of rocks in many mountains of the world (Agarwal et al. 2007). The components of MP are as follows: gums, albuminoids, calcium, potassium, nitrogen, silica, resin, vegetable matter, magnesium, sulfur, iron, chloride, phosphorous, iodine, glycoside, tannic acid, benzoic acid and several vitamins, enzymes and carrier molecules including dibenzo-a-pyrones along with humic, and fulvic acids (Ghosal 1990; Park et al. 2006). In addition to the traditional use of MP, preclinical researches have also been done on its activities, and its therapeutic effects such as antiinflammatory and antioxidant roles, antiviral and antifungal activities, analgesic effect, and protection of mast cells from degranulation have been identified (Acharya et al. 1988; Ghosal et al. 1989, 1995; Goel et al. 1990; Bhattacharya et al. 1995; Rachana 2009).
Previous studies have shown that infusion of several types of cells into the body quickly results in cell death; or after intravenous injection, MSCs reach the target organ at low or very low frequencies (Silva et al. 2008; Kang et al. 2012). On the other hand, for some reason, such as rapid clearance of factor from application site or low half-life of them in circulation, systematical or local application of exogenous factor is not a suitable option (Liu et al. 1994; Moreno et al. 2005; Ohno et al. 2007). Tissue engineering is a multidisciplinary field that is designed to repair lost or damaged tissue through the use of cells, extracellular matrix (ECM), and signals (Gholipour-Kanani et al. 2012). With the help of tissue engineering, cells and growth factors can be embedded in a scaffold and then implanted in the desired anatomic region as a wound dressing or prosthesis (De La Puente et al. 2011). The best wound dressing should be able to help regenerate damaged tissue by protecting the wound tissue from infection, reducing inflammation, and inducing cell proliferation (Kulac et al. 2013). In the present study, to reduce inflammation in burn injury, we innovatively used the acellular ovine small intestine submucosa (AOSIS) as the carrier for MP and Wharton’s Jelly Mesenchymal Stem Cells (WJ-MSCs) and modified it through MP loading and WJ-MSCs seeding. To investigate the role of modified AOSIS in inflammation, the aims of this p were to: (1) determine the loading and releasing of MP quantitatively, (2) evaluate the inflammatory effects of modified AOSIS in the burn model.

Authors: Dr.Leila Roshangar

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