Behavior and biocompatibility of rabbit bone marrow mesenchymal stem cells with bacterial cellulose membrane

Abstract

BackgroundTissue engineering has been shown to exhibit great potential for the creation of biomaterials capable of developing into functional tissues. Cellular expansion and integration depends on the quality and surface-determinant factors of the scaffold, which are required for successful biological implants. The objective of this research was to characterize and evaluate the in vitro characteristics of rabbit bone marrow mesenchymal stem cells (BM-MSCs) associated with a bacterial cellulose membrane (BCM). We assessed the adhesion, expansion, and integration of the biomaterial as well as its ability to induce macrophage activation. Finally, we evaluated the cytotoxicity and toxicity of the BCM.MethodsSamples of rabbit bone marrow were collected. Mesenchymal stem cells were isolated from medullary aspirates to establish fibroblast colony-forming unit assay. Osteogenic, chondrogenic, and adipogenic differentiation was performed. Integration with the BCM was assessed by scanning electron microscopy at 1, 7, and 14 days. Cytotoxicity was assessed via the production of nitric oxide, and BCM toxicity was assessed with the MTT assay; phagocytic activity was also determined.ResultsThe fibroblastoid colony-forming unit (CFU-F) assay showed cells with a fibroblastoid morphology organized into colonies, and distributed across the culture area surface. In the growth curve, two distinct phases, lag and log phase, were observed at 15 days. Multipotentiality of the cells was evident after induction of osteogenic, chondrogenic, and adipogenic lineages. Regarding the BM-MSCs’ bioelectrical integration with the BCM, BM-MSCs were anchored in the BCM in the first 24 h. On day 7 of culture, the cytoplasm was scattered, and on day 14, the cells were fully integrated with the biomaterial. We also observed significant macrophage activation; analysis of the MTT assay and the concentration of nitric oxide revealed no cytotoxicity of the biomaterial.ConclusionThe BCM allowed the expansion and biointegration of bone marrow progenitor cells with a stable cytotoxic profile, thus presenting itself as a biomaterial with potential for tissue engineering.

Bacterial cellulose: a versatile biopolymer for wound dressing applications

“…Another example used rabbit bone marrow MSC (BM‐MSC) associated with BC (BC/BM‐MSC). The BM‐MSC were observed, by scanning electron microscopy (SEM), to have fully integrated within the BC matrix and to have the ability to differentiate into more than one mesenchymal lineage (chondrogenic, osteogenic or adipogenic) once integrated into the matrix, yielding membranes with good biocompatibility results (Silva et al ., ). Besides stem cells, other types of cells were added to BC membranes; human epidermal keratinocytes and dermal fibroblasts (DF) were incorporated in a BC/acrylic acid (AA) hydrogel exhibiting wound healing ability in vitro and in in vivo models (Fig.…”

“…In the same study, cytotoxicity was also assessed through the measurement of nitric oxide, produced by macrophages to eliminate pathogens, as an inflammatory response mediator, inhibiting or inducing inflammation. The colorimetric read of the NO released in the presence of the BC showed a non‐cytotoxic concentration (Silva et al ., ).…”

Scaffolds for Chondrogenic Cells Cultivation Prepared from Bacterial Cellulose with Relaxed Fibers Structure Induced Genetically

“…In the both cited studies, the electrospun nano-sized fibers were less supportive for chondrogenesis than micro-sized ones. Importantly, even though many reports indicated the stimulation of ECM secretion from chondrocytes by BNC scaffolds both in vitro and in vivo [29,30,31,32,33,34,35], our material is the first one modified by means of genetic engineering and showing an increasing GAG secretion when compared to the native BNC (Figure 5b). The more so because most advances in the BNC usage as scaffold material in tissue engineering has been achieved by chemical and physical modifications, thus bringing the risk of impairing natural biocompatibility properties of BNC [11,12,20,21,29,32,33,34].…”

“…These cells cultivated in vitro on the plastic surfaces dedifferentiate quickly to less specific cells with enhanced ability to proliferate. Morphological changes following chondroneogenesis pathway can be clearly distinguished as cells tend to form nodules (groups of cells connected with each other) and are rounded in shape (in contrast to stretched, elongated fibrous-like cells) [35]. The second approach to verify and quantify the advancement of chondrogenic differentiation is estimation of glycosaminoglycans (an important ECM component) synthesis level, most commonly by staining with Alcian blue [10].…”

“…Most of the progress in the field of development of chondrogenic cells supporting material based on bacterial cellulose included chemical or physical modifications, disturbing the natural form of BNC. Nevertheless, good integration of native bacterial cellulose membranes with mammalian cells has been shown constantly, with a recent example of rabbit bone marrow mesenchymal stem cells differentiating on this material into three lineages: chondrogenic, osteogenic, and adipogenic [35].…”