A hybrid hydrogel was obtained from decellularized extract from Wharton’s jelly (DEWJ) and silk fibroin (SF) and characterized for cartilage tissue engineering. Wharton’s jelly was used due to its similarity with articular cartilage in extracellular matrix composition. Also, silk fibroin has good mechanical properties which make this construct appropriate for cartilage repair. Decellularization of Wharton’s jelly was verified by DAPI staining, DNA quantification, and PCR analysis. Then, the biochemical composition of DEWJ was determined by ELISA kits for total proteins, collagens, sulfated glycosaminoglycans (sGAG), and transforming growth factor β1 (TGF-β1). After fabricating pure SF and SF/DEWJ hybrid hydrogels, their physical and mechanical properties were characterized by FESEM, Fourier-transform infrared spectroscopy (FTIR) and rheological assays (amplitude and frequency sweeps). Furthermore, cell viability and proliferation were assessed by MTT assay. The results have shown that DEWJ in hybrid hydrogels enhances mechanical properties of the construct relative to pure SF hydrogels. Also, this extract at its 40% concentration in culture media and 20% or 40% concentrations in SF/DEWJ hybrid hydrogels significantly increases population of the cells compared to control and pure SF hydrogel after 7 days. In conclusion, this study proposes the potential of SF/DEWJ hybrid hydrogels for cartilage tissue engineering applications.
Extracellular matrix-based scaffolds derived from mammalian tissues have been used in tissue engineering applications. Among all the tissues, decellularized small intestine submucosal layer (SIS) has been recently investigated for its exceptional characteristics and biocompatibilities. These investigations have been mainly focused on the decellularized porcine SIS; however, there has not been any report on ovine SIS (OSIS) layer. In this study, OSIS was decellularized and its physical, chemical, and morphological properties were evaluated. Decellularization was carried out using chemical reagents and various physical conditions. The effects of different conditions were evaluated on histological and biomechanical properties, quality of residual DNA, GAPDH gene expression, and biocompatibility. Results revealed satisfactory decellularization of OSIS which could be due to its thin thickness. Mechanical properties, structural form, and glycosaminoglycan contents were preserved in all the decellularized groups. In SDS-treated groups, further cells and DNA residues were removed compared to the groups treated with Triton X-100 only. No toxicity was observed in all treatments, and viability, expansion, and cell proliferation were supported. In conclusion, our results suggest that OSIS decellularized scaffold could be considered as an appropriate biological scaffold for tissue engineering applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 933-944, 2018.
A remarkable challenge in myocardial tissue engineering is the development of biomimetic constructs that can potentially improve myocardial repair and regeneration. Polyurethane (PU) scaffolds are extensively utilized in the cardiovascular system. We have synthesized a new biodegradable poly(ester-ether urethane urea) (PEEUU) using a new and simple method. To enhance mechanical and physicochemical properties, the PEEUU was blended with polycaprolactone (PCL). We then fabricated a series of new PU–PCL scaffolds. The scaffolds were then characterized using SEM, porosity measurement, attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), DSC, water contact angle measurement, swelling measurement, in vitro degradation rate, and mechanical tests. Expression of the cardiac-specific proteins on the scaffolds was investigated using immunofluorescence staining and quantitative real-time PCR. The elasticity of blends increased with an increase of PEEUU. In the blend scaffolds, the size and interconnectivity of pores were in an appropriate range (142–170 μm) as reported in the literature. These blend scaffolds revealed high cell metabolic activity for cardiomyoblasts and also enabled cells to proliferate and express cardiac marker proteins at higher rates. Histological examination of subcutaneously transplanted scaffolds after two months revealed degradation in the blend scaffolds. It is demonstrated that functionality of cells is sensitive to the composition of biomaterials used, and the effective cell–biomaterial interactions are critical in order to create a functional tissue engineered product that allows seeded cells to develop their normal activity. The PEEUU–PCL blends could potentially provide a versatile platform to fabricate functional scaffolds with an effective cell–biomaterial interaction for cardiac tissue regeneration.
Decellularized extracellular matrices (ECM) based materials are routinely used for a variety of clinical applications. Hereof, in vivo application of decellularized ovine small intestinal submucosal (DOSIS) layer as, a scaffold is yet to be investigated. In this study, the effectiveness of the DOSIS scaffold, with or without rat bone marrow mesenchymal stem cells (BM-MSCs), in full-thickness wound healing of critical-sized defect was experimentally studied in a rat model. The experimental groups included; group I (control), group II (DOSIS), and group III (BM-MSCs-seeded DOSIS). Wound healing of all groups was examined and compared clinically and histopathologically on days 7, 14, and 21 postoperation. Our results represented BM-MSCs-seeded DOSIS accelerated wound contraction and healing compared to both the DOSIS alone and control groups. Epithelization was close to completion 21 days postoperation in DOSIS alone. In OSIS with BM-MSCs group, epithelization was faster and had fully taken place at the subsequent time points. DOSIS layer, as cell-free form with low substantially DNA content, accelerated healing of rat skin wound defects that was created at critical-size and full-thickness. In conclusion, decellularized OSIS alone and in combination with BM-MSCs has the potential to be used as a wound graft material in skin regenerative medicine. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2177-2190, 2018.
The extracellular matrix of different mammalian tissues is commonly used as scaffolds in the field of tissue engineering. One of these tissues, which has frequently been studied due to its structural and biological features, is the small intestine submucosal membrane.These research are mainly done on the porcine small intestine. However, a report has recently been published about a scaffold produced from the submucosal layer of the ovine small intestine. In the present study, ovine small intestine submucosal (OSIS) was decellularized in a modified manner and its histological, morphological, and biomechanical properties were studied. Decellularization was performed in two phases: physical and chemical. In this method, a chloroform-methanol mixture, enzymatic digestion, and a constant dose of sodium dodecyl sulfate (SDS) was used in the least agitation time and its histological property and biocompatibility were evaluated in the presence of adipose tissue-derived stem cells (ADSCs); furthermore, ADSCs were isolated with a simple method (modified physical washing non-enzymatic isolation). The results were showed that the use of OSIS could be effective and operative. Mechanical properties, histological structure and shape, and glycosaminoglycan content were preserved. In the SDS-treated group, more than 90% of the native cells of tissue were deleted, and also in this group, no toxicity was observed and cell proliferation was supported, compared to the untreated group. Therefore, our results indicate that ADSCs seeded on OSIS scaffold could be used as a new approach in regenerative medicine as hybrid or hydrogel application.
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