Abstract
The porous composite scaffolds (PHBV/HA) consisting of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and hydroxyapatite (HA) were fabricated using a hot-press machine and salt-leaching. Collagen (type I) was then immobilized on the surface of the porous PHBV/HA composite scaffolds to improve tissue compatibility. The structure and morphology of the collagen-immobilized composite scaffolds (PHBV/HA/Col) were investigated using a scanning electron microscope (SEM), Fourier transform infrared (FTIR), and electron spectroscopy for chemical analysis (ESCA). The potential of the porous PHBV/HA/Col composite scaffolds for use as a bone scaffold was assessed by an experiment with osteoblast cells (MC3T3-E1) in terms of cell adhesion, proliferation, and differentiation. The results showed that the PHBV/HA/Col composite scaffolds possess better cell adhesion and significantly higher proliferation and differentiation than the PHBV/HA composite scaffolds and the PHBV scaffolds. These results suggest that the PHBV/HA/Col composite scaffolds have a high potential for use in the field of bone regeneration and tissue engineering.
Highlights
Polyhydroxyalkanoates (PHAs) are natural biodegradable thermoplastics that are accumulated by a wide variety of microorganisms as a unique intracellular storage of carbon, among which poly (3-hydroxybutyric acid) (PHB) and poly (3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) are two main products [1]
After 15 days of culturing the osteoblasts on the PHBV, PHBV and HA (PHBV/HA) and PHBV/HA/Col nanofibrous scaffolds, alkaline phosphatase (ALP) staining was done by a standard procedure [30] according to the manufacturer’s instructions (Alkaline phosphatase, Leukocyte, Procedure no. 86, Sigma-Aldrich, USA)
Since ALP is one of the most exclusive proteins synthesized by osteoblasts, the presence of ALP in MC3T3-E1 osteoblasts which were cultured on PHBV/HA/Col composite scaffolds could be used as the sole marker to confirm the osteoblastic phenotype of the cells [36]
Summary
Polyhydroxyalkanoates (PHAs) are natural biodegradable thermoplastics that are accumulated by a wide variety of microorganisms as a unique intracellular storage of carbon, among which poly (3-hydroxybutyric acid) (PHB) and poly (3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) are two main products [1]. To mimic the natural environment of connective tissue, the polymeric surfaces have been modified by coating or grafting ECM proteins (fibronectin, vitronectin, and collagen) that have a cellbinding domain containing the RGD sequence [15,16,17,18,19]. Depending on the final application (cell carrier/scaffold), the natural protein is either grafted (chemically immobilized) or dip coated (physically immobilized) on the polymer surface [14, 21]. The combination of PHBV, HA, and collagen in a porous scaffold should give the combined benefits of properties which are not achievable by individual components. Cell behaviors on PHBV/HA/Col, PHBV/HA, and PHBV scaffolds were compared, in terms of cell adhesion, proliferation, and differentiation to study the potential for use in the field of bone regeneration. (c) Figure 1: FE-SEM images of porous PHBV (a), PHBV/HA (b), and PHBV/HA/Col (c) scaffolds
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