Development and evaluation of a novel beneficent antimicrobial bioscaffold based on animal waste-fish swim bladder (FSB) doped with silver nanoparticles.

Treated fish wastes have found many applications in industry and medicine. Besides, nowadays low-cost scaffold with antimicrobial activity which can accelerates the process of wound healing is very demanding. In this study fish swim bladder (FSB), taken from Rutilus frisii, which is a disposable waste was doped with silver nanoparticles (AgNPs) and evaluated as antimicrobial wound dressing. The scanning electron microscopy (SEM) micrographs showed the presence of AgNPs on the scaffold. Histological observation confirmed cells and muscle removal from FSB and collagen preservation. There was significant antibacterial activity even in 50 ppm AgNPs concentration against pathogenic bacteria, swelling ratio was rather low, and cytotoxic assay revealed that the AgNPs-FSB scaffold had no toxic effect on human foreskin fibroblast (HFF) cells. Interestingly, despite the porous structure, the AgNPs-FSB scaffold was found to be a suitable barrier to microbial penetration even after 72 h. Further study showed the gradual release of AgNPs during 24 h. In conclusion, biofabricated FSB prepared in this study have appropriate characteristics notably encompassing a high quantity of collagen and broad-spectrum antimicrobial activity. Also, its porous structure made it suitable as a 3-D structure for the growth of cells and adding other antimicrobial nano-sized materials.

Effect of swim bladder matrix treated with hyaluronic acid on wound healing: an animal model evaluation.

OBJECTIVE: To investigate the potential restorative effects of the white fish (Rutilus frisii kutum) swim bladder matrix (FSBM) treated with hyaluronic acid (HA) in the repair of skin wounds in an animal model. METHOD: Wistar rats were used to investigate the repair function of FSBM as a biological scaffold. Rats were given four round wounds on their back. Wounds were divided into four groups: control, acellular fish swim bladder matrix (AFSBM), HA and AFSBM impregnated with HA (AFSBM-HA). Macroscopic and histological studies in wound healing were investigated on days three, five and seven after injury. RESULTS: The AFSBM-HA group showed a rapid reduction in wound area compared with the other groups, indicating faster wound healing (p<0.001). At day seven after injury, the AFSBM group showed a faster process in epidermal layer formation and angiogenesis compared with the control group (p<0.001). In the AFSBM-HA group, in addition to the reduction of inflammatory cells density, a significant increase in the number of fibroblast cells and collagen synthesis was observed. An elevation in collagen synthesis and angiogenesis was also observed in the HA group compared with the control group. CONCLUSION: The findings of this study indicate that AFSBM-HA can potentially be used as a biological dressing in wound healing.

The in vitro analysis of migration and polarity of blastema cells in the extracellular matrix derived from bovine mesenteric in the presence of fibronectin.

Cell migration is an essential process in embryonic development, wound healing, and pathological conditions. Our knowledge of cell migration is often based on the two dimentional evaluation of cell movement, which usually differs from what occurred in vivo. In this study, we investigated cellular migration from blastema tissue toward bovine decellularized mesentery tissue. In this regard, fibronectin (FN) was assessed to confirm cell migration. Therefore, we established a cell migration model using blastema cells migration toward the extracellular matrix derived from bovine mesenteric tissue. A physiochemical decellularization method was utilized based on freeze-thaw cycles and agitation in sodium dodecyl sulfate and Triton X-100 to remove cells from the extracellular matrix (ECM) of bovine mesenteric tissue. These types of matrices were assembled by the rings of blastema tissues originated from the of New Zealand rabbits pinna and cultured in a medium containing FN in different days in vitro, and then they are histologically evaluated, and the expression of the Tenascin C gene is analyzed. By means of tissue staining and after confirmation of the cell removal from mesenteric tissue, polarity, and migration of blastema cells was observed in the interaction site with this matrix. Also, the expression of the Tenascin C gene was assessed on days 15 and 21 following the cell culture process. The results showed that the three dimentional model of cellular migration of blastema cells along with the ECM could be a suitable model for investigating cell behaviors, such as polarity and cell migration in vitro.

Decellularized kidney in the presence of chondroitin sulfate as a natural 3D scaffold for stem cells.

OBJECTIVES: Use of biological scaffolds and automating the cells directing process with materials such as growth factors and glycosaminoglycans (GAGs) in a certain path may have beneficial effects in tissue engineering and regenerative medicine in future. In this research, chondroitin sulfate sodium was used for impregnation of the scaffolds. It is a critical component in extracellular matrix and plays an important role in signaling pathway; however, little is known about its role within mammalian development and cell linage specification. MATERIALS AND METHODS: Due to its porous and appropriate structure and for putting cells in 3D space, the kidney of BALB/c mouse was selected and decellulalized using physical and chemical methods. After decellularization, the scaffold was impregnated in chondroitin sulfate solution (CS) for 24 hr. Then, 60×10(5) human adipose-derived mesenchymal stem cells were seeded on the scaffold to assess their behavior on day 5, 10, 15, 20, and 25. RESULTS: After 48 hr, DAPI staining approved completed decellularized kidney by 1% SDS (sodium dodecyl sulfate). Migration and establishment of a number of cells to the remaining area of the glomerulus was observed. In addition, cell accumulation on the scaffold surface as well as cells migration to the depth of kidney formed an epithelium-like structure. Up to the day 15, microscopic study of different days of seeding showed the gradual adhesion of large number of cells to the scaffold. CONCLUSION: Glycosaminoglycan could be a right option for impregnation. It is used for smartification and strengthening of natural scaffolds and induction of some behaviors in stem cells.

Decellularized Bovine Articular Cartilage Matrix Reinforced by Carboxylated-SWCNT for Tissue Engineering Application

ABSTRACT Nanotubes with their unique properties have diversified mechanical and biological applications. Due to similarity of dimensions with extracellular matrix (ECM) elements, these materials are used in designing scaffolds. In this research, Carboxylated Single-Wall Carbon Nanotubes in optimization of decellularized scaffold of bovine articular cartilage was used. At first, the articular cartilage was decellularized. Then the scaffolds were analyzed in: (i) decellularized scaffolds, and (ii) scaffolds plunged into homogenous suspension of nanotubes in distilled water, were smeared with Carboxylated-SWCNT. The tissue rings derived from the rabbit's ear were assembled with reinforced scaffolds and they were placed in a culture media for 15 days. The scaffolds in two groups and the assembled scaffolds underwent histologic and electron microscopy. Scanning electron microscopy showed that the structure of ECM of articular cartilage has been maintained well after decellularization. Fourier transform infrared analysis showed that the contents of ECM have not been changed under treatment process. Atomic force microscopy analysis showed the difference in surface topography and roughness of group (ii) scaffolds in comparison with group (i). Transmission electron microscopy studies showed the Carboxylated-SWCNT bond with the surface of decellularized scaffold and no penetration of these compounds into the scaffold. The porosity percentage with median rate of 91.04 in group (i) scaffolds did not have significant difference with group (ii) scaffolds. The electron microscopy observations confirmed migration and penetration of the blastema cells into the group (ii) assembled scaffolds. This research presents a technique for provision of nanocomposite scaffolds for cartilage engineering applications.