The current study aimed to assess the microbial quality of municipal (tap) and ground (borehole) water in Karachi, Pakistan. A health survey was also conducted to assess possible health risks of the drinking water. Fifty water samples (n = 25 each of tap and ground water) were collected from various locations of five administrative districts of Karachi for bacteriological analysis. In addition, a survey was conducted to assess the impact of drinking water on the health of city residents. Microbiological analysis results showed the presence of total coliform in 48 out of 50 (96%) tested samples. The total viable plate count at 37 °C was >200 CFU/ml in the majority of the collected samples which exceeded the permissible limit set by the World Health Organization (WHO) and the Pakistan Environmental Protection Agency. To evaluate the health risk of contaminated water, a total of 744 residents were interviewed. The information acquired from this field work revealed a high prevalence of waterborne diseases in the order of diarrhea and vomiting > skin problems > malaria > prolonged fever > eye problems and jaundice. To solve water and environmental problems, awareness and regular monitoring programs of water management and safe disposal of waste have been suggested.
This study was aimed to enhance the healing potential of rat bone marrow mesenchymal stem cells against chronic diabetic wounds through interleukin-7 (IL-7) transfection. IL-7 plays an important role in wound healing and acts as a survival factor in some cell types. This study involves isolation, propagation, and characterization of mesenchymal stem cells (MSCs) and their modification with IL-7 gene via retroviral transfection. Transfected MSCs were assessed for their effect on angiogenic genes by qPCR. Wound healing potential of transfected MSCs was analyzed by scratch assay in vitro and by transplanting these cells in rat diabetic wound models in vivo. Wound area was measured for a period of 15 days and subsequent histological analysis was performed. qPCR results showed increased expression of IL-7 gene (p ≤ 0.05) and also principal angiogenic genes, vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), VEGF receptor 1 (FLT-1), and VEGF receptor 2 (FLK-1) (p ≤ 0.05). Neuropilin-1 (NRP-1) did not show any significant change. In vitro analysis of IL-7 MSCs showed intense cell-cell connections and tube formation as compared to the normal MSCs. Rate of wound closure was more (p ≤ 0.001) in case of diabetic group transplanted with IL-7 MSCs. Histological examination revealed enhanced vascular supply in skin tissues of diabetic animals transplanted with IL-7 transfected MSCs as compared to normal MSCs. Immunohistochemical results showed significantly higher expression of IL-7 (p ≤ 0.001) and α-smooth muscle actin(p ≤ 0.001) in the tissue sections of IL-7 transfected group as compared to normal MSCs and the diabetic control group; the latter indicates increase in the number of blood vessels. It is concluded from this study that IL-7 overexpression in MSCs can enhance the healing potential of MSCs and aid in wound closure in diabetic animals through the induction of angiogenic genes.
Hypoxic wounds are tough to heal and are associated with chronicity, causing major healthcare burden. Available treatment options offer only limited success for accelerated and scarless healing. Traditional skin substitutes are widely used to improve wound healing, however, they lack proper vascularization. Mesenchymal stem cells (MSCs) offer improved wound healing; however, their poor retention, survival and adherence at the wound site negatively affect their therapeutic potential. The aim of this study is to enhance skin regeneration in a rat model of full-thickness dermal wound by transplanting genetically modified MSCs seeded on a three-dimensional collagen scaffold. Rat bone marrow MSCs were efficiently incorporated in the acellular collagen scaffold. Skin tissues with transplanted subcutaneous scaffolds were histologically analysed, while angiogenesis was assessed both at gene and protein levels. Our findings demonstrated that three-dimensional collagen scaffolds play a potential role in the survival and adherence of stem cells at the wound site, while modification of MSCs with jagged one gene provides a conducive environment for wound regeneration with improved proliferation, reduced inflammation and enhanced vasculogenesis. The results of this study represent an advanced targeted approach having the potential to be translated in clinical settings for targeted personalized therapy.
Background: Time-dependent initiation of wound healing phases and their associated healing mediators are crucial for injured skin regeneration. Mesenchymal stem cells (MSCs) secrete various paracrine factors which aid in wound healing via acceleration of cell migration, angiogenesis, tissue granulation, and modulation of inflammation at the wound site.
Objective: This study was aimed to investigate thetherapeutic effect of human umbilical cord MSCs (hUCMSCs) in the regeneration of cold-induced burn wound model.
Methods: hUCMSCs were characterized by immunocytochemistry and flow cytometry. Scratch assay was performed using rat skin fibroblasts treated with conditioned medium of hUCMSCs. An in vivo cold burn wound model was developed and hUCMSCs were locally transplanted. Macroscopic analysis of wound closure was done at days 1, 3, 7 and 14 corresponding to wound healing phases. Gene expression, histology and immunohistochemical analysis were performed to confirm complete wound repair.
Results: We observed a significant reduction in the scratch area in the treated group as compared to the control. Wound area was remarkably reduced in the burn wound model transplanted with hUCMSCs well before the end of the experimental period (day 14). Histology showed intact collagen with regenerated epidermis, dermis and hair follicles, while immunohistochemistry showed enhanced angiogenesis in the last phase of healing in the treated group. Temporal gene expression showed significant reduction in inflammatory cytokines and upregulation of pro/angiogenic and remodeling cytokines at particular time points.
Conclusion: It is concluded from this study that hUCMSCs accelerate wound closure with enhanced neovascularization and reduced inflammation in rat dermal wounds.
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