RESUMO
Osteochondral tissue is hard to regenerate after injuries or degenerative diseases. Traditional treatments still have disadvantages, such as donor tissue availability, donor site morbidity, implant loss, and limited durability of prosthetics. Thus, recent studies have focused on tissue engineering strategies to regenerate osteochondral defects with different scaffold designs. Scaffolds have been developed from monolayer structures to bilayer scaffolds to repair the cartilage-bone interface and to support each tissue separately. In this study, Si-substituted nanohydroxyapatite particles (Si-nHap) and silica-based POSS nanocages were used as reinforcements in different polymer layers to mimic a cartilage-bone tissue interface. Chitosan and zein, which are widely used biopolymers, are used as polymer layers to mimic the structure. This study reports the development of a bilayer scaffold produced via fabrication of two different nanocomposite layers with different polymer-inorganic composites in order to satisfy the complex and diverse regenerative requirements of osteochondral tissue. The chitosan/Si-nHap microporous layer and the zein/POSS nanofiber layer were designed to mimic a bone-cartilage tissue interface. Bilayer scaffolds were characterized with SEM, compression, swelling, and biodegradation tests to determine morphological, physical, and mechanical properties. The results showed that the bilayer scaffold had a structure composed of microporous and nanofiber layers joined at a continuous interface with appropriate mechanical properties. Furthermore, in vitro cell culture studies have been performed with LDH, proliferation, fluorescence imaging, and ALP activity assays using osteosarcoma and chondrosarcoma cell lines. ALP expression levels provide a good illustration of the improved osteogenic potential of a porous chitosan/Si-nHap layer due to the Si-doped nHap incorporation. Histological data showed that both fiber and porous layers that mimic the cartilage and bone sections exhibit homogeneous cell distribution and matrix formation. Histochemical staining was used to determine the cell proliferation and ECM formation on each layer. In vitro studies indicated that zein-POSS/chitosan/Si-nHap nanocomposite bilayer scaffolds showed promising results for osteochondral regeneration.
RESUMO
The current study reports the biocompatibility and biomechanical characteristics of loofah-based scaffolds combined with hydroxyapatite (HA), cellulose, poly-l-lactic acid (PLLA) with chondrocytes-like cells. Scanning electron microscope (SEM) micrographs of the scaffolds showed that the addition of PLLA usually resulted in an increase in cell's attachment on scaffolds. Mechanical and elemental analyzes were assessed using tensile test and Energy Dispersive X-ray spectrometry (EDS), respectively. In summary, we showed that the loofah+PLLA+HA scaffolds perform significantly better than other loofah-based scaffolds employed in terms of increasing a diversity of mechanical properties including tensile strength and Young's modulus. Based on the analysis of the differential scanning calorimetry (DSC) thermograms and EDS spectrums that give an idea about the calcium phosphate (CaP) ratios, the improvement in the mechanical properties could principally be recognized to the strong interaction formed between loofah, PLLA and HA. The viability of chondrocytes on loofah-based scaffolds was analyzed by XTT tests. However, none of the scaffolds have proved to be toxic in metabolic activity. The histological evaluation obtained by hematoxylin and eosin (H&E), Masson trichrome, toluidine blue and immunohistochemistry methods showed that cells in all scaffolds produced extracellular matrix that defined proteoglycan and type I-II collagens. The results of this study suggest that the loofah-based scaffold with desirable properties can be considered as an ideal candidate for cartilage tissue engineering applications.
