Evaluation of a multilayered chitosan-hydroxy-apatite porous composite enriched with fibronectin or an in vitro-generated bone-like extracellular matrix on proliferation and diferentiation of osteoblasts

Abstract

The use of extracellular matrix (ECM) molecules from tissues is an interesting way to induce specific responses of cells grown onto composite scaffolds to promote adhesion, proliferation and differentiation. There have been several studies on the effects on cell proliferation and differentiation of osteoprogenitor cells cultured onto composites, either adding some ECM molecules or grown in the presence of growth factors. Other studies involve the use of osteoblasts cultured on a three-dimensional (3D) matrix, enriched with ECM molecules produced by the same cells grown previously inside the composite. Here, the effect of enrichment of a novel multilayered chitosan-hydroxyapatite composite with ECM molecules produced by osteoblasts, or the addition of 25 or 50 µg/ml fibronectin to the composite, on proliferation and differentiation of osteoblasts cultured on these composites was studied. The results showed an increase in the number of osteoblasts from day 1 of culture, which was higher in the group grown onto composites enriched with the highest concentration of fibronectin or with ECM molecules produced naturally by osteoblasts cultured previously on them, when compared with the control group. However, this increment tended to decline in all groups after day 7 of culture, the day when they reached the highest peak of proliferation. Differentiation expressed as alkaline phosphatase activity followed the proliferation pattern of the cells cultivated on the scaffolds. The results demonstrate the potential offered by these enriched 3D multilayered composites for improving their ability as bone grafting material.

Guided bone regeneration using beta‐tricalcium phosphate with and without fibronectin—An experimental study in rats

“…In recent years, extensive experimental research has focused on different approaches to accelerate bone regeneration (Gomes & Fernandes, ; Li et al, ; Luvizuto et al, ; Rodriguez et al, ). Tissue engineering using combinations of cells, scaffolds and bioactive factors is novel possibilities as bone grafts for skeletal defects (Annibali et al, ; Ball, O'Connor, & Pandit, ; Fernández et al, ). Fibronectin is a glycoprotein of the extracellular matrix that promotes cell adhesion, differentiation and expansion, and has been investigated in combination with regeneration biomaterials for improving proliferation and differentiation of osteoblasts cultivated on composite scaffolds (Ball et al, ; Fernández et al, ).…”

“…Tissue engineering using combinations of cells, scaffolds and bioactive factors is novel possibilities as bone grafts for skeletal defects (Annibali et al, ; Ball, O'Connor, & Pandit, ; Fernández et al, ). Fibronectin is a glycoprotein of the extracellular matrix that promotes cell adhesion, differentiation and expansion, and has been investigated in combination with regeneration biomaterials for improving proliferation and differentiation of osteoblasts cultivated on composite scaffolds (Ball et al, ; Fernández et al, ). Moreover, it has been shown that anodized titanium implants treated with fibroblast growth factor‐Fn fusion protein enhanced osseointegration (Park et al, ).…”

Comparative effect of human andTrypanosoma cruzicalreticulin in wound healing

“…The use of chitosan‐based scaffold biomaterials for different tissue engineering applications is well known (Fernandez et al ., ; Kim et al ., ). Because of the chitosan swelling behaviour and other biocompatible properties (Yazdani‐Pedram et al ., ), its use as an implant embedded with human or parasite CRTs was explored.…”

“…The scaffolds were prepared as follows: a solution of 2% low molecular weight chitosan (20–200 cps; Aldrich), 86% deacetylated, in 5% acetic acid was shaken for 24 h at room temperature and then filtered to remove non‐dissolved chitosan and impurities (Fernández et al ., ). Chitosan sponges were obtained by freeze‐drying the chitosan solution with the addition of 0–5% glutaraldehyde (crosslinker agent) in liquid nitrogen and then lyophilized repeatedly for 48 h. 4 mm 2 pieces, with an average weight of 20 µg, were then cut and individually packed and sterilized by EThO gas.…”

Preventing infection of osseointegrated transcutaneous implants: Incorporation of silver into preconditioned fibronectin-functionalized hydroxyapatite coatings suppresses Staphylococcus aureus colonization while promoting viable fibroblast growth in vitro

“…Studies have also shown that the addition of fibronectin improves osteoblast responses further. [59][60][61] If osteoblasts were found to respond well to HAAg/HAAgFn, this technique could be useful for the intraosseous portion of osseointegrated transcutaneous implants as well as more widely for other orthopedic implants such as arthroplasty prostheses. This would be advantageous because hip and knee arthroplasties are associated with an infection rate of approximately 1%, which is associated with significant morbidity and substantial economic costs.…”