Multilayer cellular stacks of gelatin-hydroxyapatite fiber scaffolds for bone tissue engineering.

Multilayer cellular stacks of crosslinked, electrospun 25 wt % hydroxyapatite (HA)-gelatin and pure gelatin fiber scaffolds, seeded with human fetal osteoblasts (hFOBs), were studied for up to 18 days in static and dynamic cell culture. Two types of stack models were investigated: a four-layer stack with cells seeded at the bottom surface of the first/top layer and the top surface of the fourth/bottom layer, so that the two middle layers were not seeded with cells with the aim to act as continuing conduits of culture medium and nutrients supply to the adjacent cell-populated zones; a three-layer stack with cells seeded at the bottom surface of each layer. hFOBs exhibited lower migration rate through the stack thickness for the 25 wt % HA-gelatin scaffolds as compared to the pure gelatin scaffolds, due to the small pores of the former. Hence, the regularly seeded three-layer stack maintained cell-free porous zones in all layers through which the culture medium could continuously perfuse, while good fusion was achieved at the interface of all layers via the cross-migrating cells with a preference to downwards vertical migration attributed to gravity. Dynamic cell culture conditions enhanced overall cell growth by about 6% for the regularly seeded three-layer stack. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 779-789, 2017.

Fabrication and characterisation of biomimetic, electrospun gelatin fibre scaffolds for tunica media-equivalent, tissue engineered vascular grafts.

It is increasingly recognised that biomimetic, natural polymers mimicking the extracellular matrix (ECM) have low thrombogenicity and functional motifs that regulate cell-matrix interactions, with these factors being critical for tissue engineered vascular grafts especially grafts of small diameter. Gelatin constitutes a low cost substitute of soluble collagen but gelatin scaffolds so far have shown generally low strength and suture retention strength. In this study, we have devised the fabrication of novel, electrospun, multilayer, gelatin fibre scaffolds, with controlled fibre layer orientation, and optimised gelatin crosslinking to achieve not only compliance equivalent to that of coronary artery but also for the first time strength of the wet tubular acellular scaffold (swollen with absorbed water) same as that of the tunica media of coronary artery in both circumferential and axial directions. Most importantly, for the first time for natural scaffolds and in particular gelatin, high suture retention strength was achieved in the range of 1.8-1.94 N for wet acellular scaffolds, same or better than that for fresh saphenous vein. The study presents the investigations to relate the electrospinning process parameters to the microstructural parameters of the scaffold, which are further related to the mechanical performance data of wet, crosslinked, electrospun scaffolds in both circumferential and axial tubular directions. The scaffolds exhibited excellent performance in human smooth muscle cell (SMC) proliferation, with SMCs seeded on the top surface adhering, elongating and aligning along the local fibres, migrating through the scaffold thickness and populating a transverse distance of 186 µm and 240 µm 9 days post-seeding for scaffolds of initial dry porosity of 74 and 83%, respectively.

Electrospinning of nanocomposite fibrillar tubular and flat scaffolds with controlled fiber orientation.

Electrospinning was used in innovative electrospinning rigs to obtain tubular and flat fibrous structures with controlled fiber orientation with the aim to be used as scaffolds for biomedical applications, more specifically in the tissue engineering of vascular and orthopedic grafts. Gelatine and hydroxyapatite (HA)-gelatine solutions of various compositions were tried and electrospinning of continuous fibers was maintained for gelatine and up to 0.30 g/g HA-gelatine solutions in 2,2,2-trifluoroethanol (TFE). Small diameter tubular scaffolds were electrospun with axial fiber orientation and flat scaffolds were cut from fiber mats electrospun around a wired drum substrate. The fibrous mats were crosslinked using a glutaraldehyde solution and subjected to image analysis of SEM micrographs, water swelling tests, and mechanical testing. Fiber diameter in the electrospun scaffolds could be varied depending on the feed solution concentration and composition whereas fiber orientation was affected by the processing conditions. After crosslinking, the 0.30 g/g HA-gelatine scaffolds absorbed the minimum amount of water after 48 h soaking and they had the highest Young's modulus, 60 MPa, and highest strength, 3.9 MPa.

Structural hierarchy of biomimetic materials for tissue engineered vascular and orthopedic grafts.

Gelatine gels and gelatine/elastin gels have been prepared to be used in tissue engineered vascular grafts. Optical microscopy and atomic force microscopy (AFM) revealed that the gelatine formed nanofibrils as in soft collagen tissues. The gelatine/elastin gels were nanocomposites with flat elastin nanodomains embedded in the gelatine matrix mimicking the structure of the tunica media in arteries. Gelatine/"hydroxyapatite" (HA) nanocomposites were prepared with the in situ production of "HA" in solution. AFM revealed "HA" solid nanoparticles of about 20 nm size embedded in the gelatine matrix, which formed a hierarchical structure similar to that of the collagen matrix in bone. The application of amagnetic field of 9.4 T resulted in the elongation and orientation of gelatine particles and orientation of gelatine microfibrils in a direction perpendicular to that of the magnetic field. (c) 2007 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2008.