Many growth factors have been studied as additives accelerating lumbar fusion rates in different animal models. However, their low hydrolytic and thermal stability both in vitro and in vivo limits their workability and use. In the proposed work, a stabilized vasculogenic and prohealing fibroblast growth factor-2 (FGF2-STAB®) exhibiting a functional half-life in vitro at 37 °C more than 20 days was applied for lumbar fusion in combination with a bioresorbable scaffold on porcine models. An experimental animal study was designed to investigate the intervertebral fusion efficiency and safety of a bioresorbable ceramic/biopolymer hybrid implant enriched with FGF2-STAB® in comparison with a tricortical bone autograft used as a gold standard. Twenty-four experimental pigs underwent L2/3 discectomy with implantation of either the tricortical iliac crest bone autograft or the bioresorbable hybrid implant (BHI) followed by lateral intervertebral fixation. The quality of spinal fusion was assessed by micro-computed tomography (micro-CT), biomechanical testing, and histological examination at both 8 and 16 weeks after the surgery. While 8 weeks after implantation, micro-CT analysis demonstrated similar fusion quality in both groups, in contrast, spines with BHI involving inorganic hydroxyapatite and tricalcium phosphate along with organic collagen, oxidized cellulose, and FGF2- STAB® showed a significant increase in a fusion quality in comparison to the autograft group 16 weeks post-surgery (p = 0.023). Biomechanical testing revealed significantly higher stiffness of spines treated with the bioresorbable hybrid implant group compared to the autograft group (p < 0.05). Whilst histomorphological evaluation showed significant progression of new bone formation in the BHI group besides non-union and fibrocartilage tissue formed in the autograft group. Significant osteoinductive effects of BHI based on bioceramics, collagen, oxidized cellulose, and FGF2-STAB® could improve outcomes in spinal fusion surgery and bone tissue regeneration.

• Publikační typ
• časopisecké články MeSH

This study presents an X-ray computed nanotomography (nano-CT) based, high-resolution imaging technique. Thanks to a voxel resolution of 540 nm, this novel technique is suitable for observing the 3D morphology of soft biopolymeric scaffolds seeded with stem cells. A sample of highly porous collagen scaffold seeded with contrasted mesenchymal stem cells (MSC) was investigated by using lab-based nano-CT. The whole volume of the sample was analysed without its destruction. To evaluate the potential of nano-CT, a comparison measurement was done using a standard microscopy technique. Scanning electron microscopy (SEM) combined with energy dispersive X-ray analysis (EDX) established an extension and local accumulation of the contrasting agent - heavy metallic osmium tetroxide. The presented imaging technique is novel as it will help to understand better the behaviour of cells while interacting with three-dimensional biomaterials. This is crucial for both experimental and clinical tissue engineering applications in order to limit the risk of uncontrolled cell growth, and potentially tumour formation. LAY DESCRIPTION: Biomaterials play a crucial role in tissue engineering by serving as 3D scaffolds for cellular attachment, proliferation, and in growth ultimately leading to new tissue formation. Cell morphology and proliferation inside the 3D scaffold are necessary to know for assessing cell viability. However, these studies are usually negatively affected by the limitations of imaging techniques. We demonstrate that X-ray computed nanotomography (nano-CT), based on high-resolution imaging technique providing voxel resolution of 540 nm, is a suitable method for observing the 3D morphology of soft biopolymeric scaffolds seeded with stem cells. A sample of highly porous collagen scaffold seeded with contrasted mesenchymal stem cells (MSC) was investigated by using a lab-based nano-CT. The whole volume of the sample was analysed without its destruction. To evaluate the potential of nano-CT, a comparison measurement was done using a standard microscopy technique. Scanning electron microscopy in a combination with energy dispersive X-ray analysis established an extension and local accumulation of the contrasting agent - heavy metallic osmium tetroxide. The presented imaging technique is novel as it will help to understand better the behaviour of cells while interacting with three-dimensional biomaterials. This is crucial for both experimental and clinical tissue engineering applications in order to limit the risk of uncontrolled cell growth, and potentially tumour formation.

• MeSH
• biokompatibilní materiály MeSH
• buněčné kultury MeSH
• kolagen chemie   MeSH
• králíci MeSH
• kultivované buňky MeSH
• mezenchymální kmenové buňky ultrastruktura   MeSH
• mikroskopie elektronová rastrovací metody   MeSH
• poréznost MeSH
• rentgenová mikrotomografie metody   MeSH
• tkáňové podpůrné struktury * MeSH
• viabilita buněk MeSH
• zobrazování trojrozměrné metody   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH