Prefabrication and free transfer of a tissue engineered composite flap – An experimental model in the rat

BACKGROUND: The technique of flap-prefabrication has been successfully established in tissue engineering: missing intrinsic vascularisation of engineered tissue can be generated in vivo by microsurgical vesselloop construction. It is possible to move engineered tissue into a defect with microsurgery. In the literature, the combination of engineered tissue covered with skin is not widely reported. OBJECTIVE: Aim of this study was to establish a model to investigate scaffold prefabrication with full thickness skin graft coverage with subsequent free tissue transfer. METHODS: 8Wistar rats were operated in 2 separate steps: 1) after creating an arteriovenous loop with the femoral vessels, a porous scaffold was placed on the loop and covered with an inguinally based skin flap. A control was implanted without loop into the contralateral groin. 2) 6 weeks later the prefabricated composite flaps were microsurgically transferred to the cervical region. Skin-island monitoring was performed with Laser Doppler-scanner after the transfer. RESULTS: Continuous loss of the skin islands was observed within 72 hours. Complications included wound-dehiscence, thrombosis and death from anaesthesia; in spite of consistent loop viability. CONCLUSION: Evaluation showed that modifications are necessary to maintain the skin-island cove

Time domain optical coherence tomography investigation of bone matrix interface in rat femurs

The materials used to fabricate scaffolds for tissue engineering are derived from synthetic polymers, mainly from the polyester family, or from natural materials (e.g., collagen and chitosan). The mechanical properties and the structural properties of these materials can be tailored by adjusting the molecular weight, the crystalline state, and the ratio of monomers in the copolymers. Quality control and adjustment of the scaffold manufacturing process are essential to achieve high standard scaffolds. Most scaffolds are made from highly crystalline polymers, which inevitably result in their opaque appearance. Their 3-D opaque structure prevents the observation of internal uneven surface structures of the scaffolds under normal optical instruments, such as the traditional light microscope. The inability to easily monitor the inner structure of scaffolds as well as the interface with the old bone poses a major challenge for tissue engineering: it impedes the precise control and adjustment of the parameters that affect the cell growth in response to various mimicked culture conditions. The aim of this paper is to investigate the interface between the femur rat bone and the new bone that is obtained using a method of tissue engineering that is based on different artificial matrixes inserted in previously artificially induced defects. For this study, 15 rats were used in conformity with ethical procedures. In all the femurs a round defect was induced by drilling with a 1 mm spherical Co-Cr surgical drill. The matrixes used were Bioss and 4bone. These materials were inserted into the induced defects. The femurs were investigated at 1 week, 1 month, 2 month and three month after the surgical procedures. The interfaces were examined using Time Domain (TD) Optical Coherence Tomography (OCT) combined with Confocal Microscopy (CM). The optical configuration uses two single mode directional couplers with a superluminiscent diode as the source centered at 1300 nm. The scanning procedure is similar to that used in any CM, where the fast scanning is en-face (line rate) and the scanning in depth is much slower (at the frame rate). The results showed open interfaces due to the insufficient healing process, as well as closed interfaces due to a new bone formation inside the defect. The conclusion of this study is that TD-OCT can act as a valuable tool in the investigation of the interface between the old bone and the one that has been newly induced due to the osteoinductive process. © 2013 SPIE

Bone regeneration assessment by optical coherence tomography and microCT synchrotron radiation

Bone grafting is a commonly performed surgical procedure to augment bone regeneration in a variety of orthopaedic and maxillofacial procedures, with autologous bone being considered as the "gold standard" bone-grafting material, as it combines all properties required in a bone-graft material: osteoinduction (bone morphogenetic proteins - BMPs - and other growth factors), osteogenesis (osteoprogenitor cells) and osteoconduction (scaffold). The problematic elements of bone regenerative materials are represented by their quality control methods, the adjustment of the initial bone regenerative material, the monitoring (noninvasive, if possible) during their osteoconduction and osteointegration period and biomedical evaluation of the new regenerated bone. One of the research directions was the interface investigation of the regenerative bone materials and their behavior at different time periods on the normal femoral rat bone. 12 rat femurs were used for this investigation. In each ones a 1 mm diameter hole were drilled and a bone grafting material was inserted in the artificial defect. The femurs were removed after one, three and six months. The defects repaired by bone grafting material were evaluated by optical coherence tomography working in Time Domain Mode at 1300 nm. Three dimensional reconstructions of the interfaces were generated. The validations of the results were evaluated by microCT. Synchrotron Radiation allows achieving high spatial resolution images to be generated with high signal-to-noise ratio. In addition, Synchrotron Radiation allows acquisition of volumes at different energies and volume subtraction to enhance contrast. Evaluation of the bone grafting material/bone interface with noninvasive methods such as optical coherence tomography could act as a valuable procedure that can be use in the future in the usual clinical techniques. The results were confirmed by microCT. Optical coherence tomography can be performed in vivo and can provide a qualitative and quantitative evaluation of the bone augmentation procedure. © OSA-2013 SPIE