Fibroin and fibroin blended three-dimensional scaffolds for rat chondrocyte culture

Abstract

BackgroundIn our previous study, we successfully developed 3-D scaffolds prepared from silk fibroin (SF), silk fibroin/collagen (SF/C) and silk fibroin/gelatin (SF/G) using a freeze drying technique. The blended construct showed superior mechanical properties to silk fibroin construct. In addition, collagen and gelatin, contain RGD sequences that could facilitate cell attachment and proliferation. Therefore, in this study, the ability of silk fibroin and blended constructs to promote cell adhesion, proliferation and production of extracellular matrix (EMC) were compared.MethodsArticular chondrocytes were isolated from rat and cultured on the prepared constructs. Then, the cell viability in SF, SF/C and SF/G scaffolds was determined by MTT assay. Cell morphology and distribution were investigated by scanning electron microscopy (SEM) and histological analysis. Moreover, the secretion of extracellular matrix (ECM) by the chondrocytes in 3-D scaffolds was assessed by immunohistochemistry.ResultsResults from MTT assay indicated that the blended SF/C and SF/G scaffolds provided a more favorable environment for chondrocytes attachment and proliferation than that of SF scaffold. In addition, scanning electron micrographs and histological images illustrated higher cell density and distribution in the SF/C and SF/G scaffolds than that in the SF scaffold. Importantly, immunohistochemistry strongly confirmed a greater production of type II collagen and aggrecan, important markers of chondrocytic phenotype, in SF blended scaffolds than that in the SF scaffold.ConclusionAddition of collagen and gelatin to SF solution not only improved the mechanical properties of the scaffolds but also provided an effective biomaterial constructs for chondrocyte growth and chondrocytic phenotype maintenance. Therefore, SF/C and SF/G showed a great potential as a desirable biomaterial for cartilage tissue engineering.

Collagen Scaffolds in Cartilage Tissue Engineering and Relevant Approaches for Future Development

“…SF also lacks cell-binding motives or other mechanism beneficial in supporting cell activities [122]. Thus, it is advantageous to blend SF with collagen [125][126][127][128][129]. Generally, collagen was blended with silk by separately adjusting each solution and subsequently mixing them before next processing (e.g., freeze-drying or electrospinning) (Fig.…”

Phase Behaviour and Miscibility Studies of Collagen/Silk Fibroin Macromolecular System in Dilute Solutions and Solid State

“…The mechanism of fibroin self-assembly during dialysis has been described by several researchers [ 17 , 18 ], and the influence of various parameters, such as concentration [ 19 ], temperature [ 20 ], and ethanol content [ 21 ] on RSF self-assembly have been analyzed. Jin et al [ 17 ] suggested a micellar structure pattern for silk fibroin chains in water in which the small hydrophilic parts of silk fibroin remain hydrated, while the large termini hydrophilic parts are located at the outer edge of micelles and the hydrophobic parts are placed between those two hydrophilic blocks [ 17 ].…”

“…Several studies on collagen/silk fibroin blends showed not only the improvement in mechanical properties of the final materials, but also a favorable environment of the mixtures for cell attachment and proliferation. Nevertheless, the blends were restricted to low collagen concentration or COL/RSF ratios, or to be able to incorporate higher collagen ratios, high temperatures are required that denature the collagen triple helix and facilitate the hydrogen bonding between the two biopolymers [ 21 , 22 , 23 , 24 , 25 , 26 , 27 ]. However, inducing electrostatic interactions between these two biopolymers, which has not been used up to now, may avoid the risk of the undesirable denaturation of collagen as the result of increasing the temperature.…”

Bombyx mori derived scaffolds and their use in cartilage regeneration: a systematic review

“…The articles studied in this systematic review highlight a variety of materials, techniques and approaches towards the goal of finding a better silk biomaterial. Silk has been used as a main component blended with several other materials in many ways: blended either in ratios with hyaluronic acid (HA) 47 , poly(L-lactic-acid) (PLLA) 48 , Chitosan 49e52 , cellulose 53 , gelatin 52 or in various percentages with chitosan and hydroxyapatite 54 , HA 55 and Fibrin/HA 59 . Chitosan and blended with SF enhances b-sheet conformation and flexibility of the blended material 56,57 .…”