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Purpose. Clinically apparent anastomotic leakage (AL) after low anterior rectal resection (LAR

Glycosaminoglycan (GAG) is a chain-like disaccharide that is linked to a polypeptide core to connect two collagen fibrils/fibers and provide the intermolecular force in a Collagen-GAG matrix which can be a valuable treatment of post-burn contractures and hypertrophic scars, which remain a challenge to reconstructive surgery. The face and neck contractures are the most difficult sites to treat. This article is meant to discuss our clinical experience in using collagen-glycosaminoglycan biodegradable matrix (Integra&reg, Integra Lifesciences Corporation, Plainsboro, NJ, USA) to reconstruct defects created by excision of contracted areas from the neck and lower face areas. Between 2009 and 2011, we had 11 patients that underwent Integra reconstructive procedures. The mean follow-up period was 18 months. For all the patients, the intake of the Integra dermal regeneration template was 100%, even if one patient developed a minor infection treated with appropriate antibiotics. The patients are very satisfied with the result. A minor problem was a small difference in skin color, but this inconvenience was compensated by good skin elasticity.

Background: Alveolar bone defects are usually the main concern when planning implant treatments for the appropriate oral rehabilitation of patients. To improve local conditions and achieve implant treatments, there are several methods used for increasing bone volume, among which one of the most successful, versatile, and effective is considered to be guided bone regeneration. The aim of this demonstrative study was to propose an innovative analysis protocol for the evaluation of the effect of photobiomodulation on the bone regeneration process, using rat calvarial defects of 5 mm in diameter, filled with xenograft, covered with collagen membrane, and then exposed to laser radiation. Methods: The animals were sacrificed at different points in time (i.e., after 14, 21, and 30 days). Samples of identical dimensions were harvested in order to compare the results obtained after different periods of healing. The analysis was performed by cross-linking the information obtained using histology and high-resolution synchrotron-based tomography on the same samples. A comparison was made with both the negative control (NC) group (with a bone defect which was left for spontaneous healing), and the positive control (PC) group (in which the bone defects were filled with xenografts and collagen membrane without receiving laser treatment). Results: We demonstrated that using photobiomodulation provides a better healing effect than when receiving only the support of the biomaterial. This effect has been evident for short times treatments, i.e., during the first 14 days after surgery. Conclusion: The proposed analysis protocol was effective in detecting the presence of higher quantities of bone volumes under remodeling after photobiomodulation with respect to the exclusive bone regeneration guided by the xenograft.

Therapeutic neoangiogenesis (TNA) holds promise as a treatment for peripheral arterial disease. Nevertheless, proper tools for in vivo pre-clinical investigation of different TNA approaches and their effects are still lacking. Here we describe a chronic ischemic hindlimb model in rats using laser Doppler quantitative evaluation of tissue perfusion. Male Wistar rats (n = 20), aged between 6−8 months, with an average weight of 287 ± 26.74 g, were used. Animals were divided into two experimental groups: group A (n = 17; hindlimb chronic ischemia model) and group B (n = 3; control). Hindlimb ischemia was induced by concomitant ligation of the right femoral and popliteal artery. Evaluation of tissue perfusion was quantified in perfusion units (PU) on a scale from 0 to 500 (500 PU = maximal detectable perfusion) by laser Doppler analysis at day 0, day 15 and day 30 after induction of ischemia. Induction of chronic ischemia in the rat hindlimb by concomitant ligation of the femoral and popliteal artery can be readily obtained but requires basic microsurgical skills. Laser Doppler analysis has shown unaltered ischemia levels throughout the study (129,17 PU ± 3.13 day 0 vs. 130,33 PU day 30 ± 3,27, p = not significant (n.s.)). We demonstrate a simple and reproducible model of chronic hindlimb ischemia in rats, with stable tissue perfusion levels that are accurately quantified using laser Doppler technology. Hence, this model can represent a valid tool for further studies involving therapeutic neoangiogenesis.

Advanced preservation injury (PI) after intestinal transplantation has deleterious short- and long-term effects and constitutes a major research topic. Logistics and costs favor rodent studies, whereas clinical translation mandates studies in larger animals or using human material. Despite diverging reports, no direct comparison between the development of intestinal PI in rats, pigs, and humans is available. We compared the development of PI in rat, porcine, and human intestines. Intestinal procurement and cold storage (CS) using histidine&ndash, tryptophan&ndash, ketoglutarate solution was performed in rats, pigs, and humans. Tissue samples were obtained after 8, 14, and 24 h of CS), and PI was assessed morphologically and at the molecular level (cleaved caspase-3, zonula occludens, claudin-3 and 4, tricellulin, occludin, cytokeratin-8) using immunohistochemistry and Western blot. Intestinal PI developed slower in pigs compared to rats and humans. Tissue injury and apoptosis were significantly higher in rats. Tight junction proteins showed quantitative and qualitative changes differing between species. Significant interspecies differences exist between rats, pigs, and humans regarding intestinal PI progression at tissue and molecular levels. These differences should be taken into account both with regards to study design and the interpretation of findings when relating them to the clinical setting.

Simple Summary Patients who have multiple associated comorbidities and need to change the aortic valve may have a contraindication to open-heart surgery, the alternative being transcatheter aortic valve implantation, which requires very precise measurements of the aortic annulus to determine the dimensions of the prostheses. Ultrasonographic imaging techniques, such as transesophageal echocardiography, are constantly evolving. The aim of our study was to compare the three-dimensional transesophageal echocardiography and multi-detector computer tomography methods, with the former being an alternative for patients who cannot undergo computer tomography because of a major contraindication. We have demonstrated that there were small differences between aortic annular measurements using multi-detector computer tomography (2.25 ± 0.19 cm) and three-dimensional transesophageal echocardiography (2.25 ± 0.15 cm). Thus, three-dimensional transesophageal echocardiography can be the solution for aortic annular measurements used to select the correct prosthesis for the transcatheter aortic valve implantation procedure in patients who cannot undergo computer tomography. Abstract Background and objectives: Transcatheter aortic valve implantation (TAVI) is a therapeutic choice for high surgical risk patients, serving as an alternative to open-heart surgery. Correct measurement of the aortic annulus, which leads to the selection of a suitable prosthesis and accurate outcome prediction, is essential for the success of TAVI. The objective of this study is to evaluate the accuracy of novel imaging te chniques in measuring the aortic annulus by comparing multi-detector computer tomography (MDCT) and three-dimensional transesophageal echocardiography (3D TEE) for the selection of the optimal prosthesis. Materials and Methods: Measurements of the aortic annulus have been performed on 25 patients using MDCT and TEE, and the correlation and agreement levels between the two measuring techniques were analyzed. MDCT measurements were used for the sizing of the prostheses. Results: MDCT and TEE measurements of aortic annular diameters were significantly correlated, with a mean difference of 0.001 cm. Conclusions: 3D TEE measurements have been in good agreement with MDCT and, therefore, 3D TEE can be used as an alternative in cases where MDCT is contraindicated or not available.

Background Rodent studies suggest that luminal solutions alleviate the mucosal injury and prolong intestinal preservation but concerns exist that excessive volumes of luminal fluid may promote tissue edema. Differences in size, structure, and metabolism between rats and humans require studies in large animals before clinical use. Methods Intestinal procurement was performed in 7 pigs. After perfusion with histidine-tryptophan-ketoglutarate (HTK), 40-cm-long segments were cut and filled with 13.5% polyethylene glycol (PEG) 3350 solution as follows: V0 (controls, none), V1 (0.5 mL/cm), V2 (1 mL/cm), V3 (1.5 mL/cm), and V4 (2 mL/cm). Tissue and luminal solutions were sampled after 8, 14, and 24 hours of cold storage (CS). Preservation injury (Chiu score), the apical membrane (ZO-1, brush-border maltase activity), and the electrolyte content in the luminal solution were studied. Results In control intestines, 8-hour CS in HTK solution resulted in minimal mucosal changes (grade 1) that progressed to significant subepithelial edema (grade 3) by 24 hours. During this time, a gradual loss in ZO-1 was recorded, whereas maltase activity remained unaltered. Moreover, variable degrees of submucosal edema were observed. Luminal introduction of high volumes (2 mL/mL) of PEG solution accelerated the development of the subepithelial edema and submucosal edema, leading to worse histology. However, ZO-1 was preserved better over time than in control intestines (no luminal solution). Maltase activity was reduced in intestines receiving luminal preservation. Luminal sodium content decreased in time and did not differ between groups. Conclusions This PEG solution protects the apical membrane and the tight-junction proteins but may favor water absorption and tissue (submucosal) edema, and luminal volumes >2 mL/cm may result in worse intestinal morphology.

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 8 Wistar 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.

Introduction.The osteoconductive materials are very important in the bone regeneration procedures. The aim of this paper is to evaluate the interface between the old bone and the new regenerated one.Materials and methods. Ten rat femurs were used for this investigation. Under strict supervision, ten holes were drilled in the rat femurs and then filled with Bio-Oss® (Geistlich, Wolhusen, Switzerland). matrix. The samples were obtained after 3 month from the procedure and were investigated by MicroCT. A common Synchrotron Radiation X-Ray micro-CT experiment was performed at the SYRMEP Beamline of the ELETTRA Synchrotron Radiation Facility (Trieste, Italy). The 900 radiographic projections were acquired with a beam energy of 29 keV over 180° (0.2 degrees step) with a resolution of 9μm. A sample – detector distance of 16 cm was considered. The tomographic reconstruction was performed by means of the common filtered back-projection method. The 3D reconstructions were obtained at the University of Ancona using specific software, VGStudio Max.Results:The synchrotron offered 900 microCT files that were used to make the 3D reconstruction of the bone. After tomographic reconstruction, 3D renderings of obtained data may be made by electronically stacking up the slices. These 3D volumes may be also sectioned in arbitrary ways, zoomed and rotated in order to locate better the individual details. While the 2D slice images and 3D renderings are very useful for making qualitative observations of an internal concrete structure, the real benefit is the quantitative information that can be extracted from the 3D datasets.The reconstructions allow observing the normal rat bone, the new regenerated bone and also the rest of nonactivated matrix of Bio-Oss® (Geistlich, Wolhusen, Switzerland). The system permits to make measurements in order to quantify the volume of the new bone formation and also the status of the bone defect, in order to define the healing process.Conclusions:The MicroCT method proves to be an important imagistic tool for nondestructive evaluation of the interface between the new regenerated bone and the old one from the rat femurs. This method allows numerical quantification of the new regenerated bone and define the healing bone process.

A high incidence of anastomotic leakage (37.5%) is reported after low anterior rectal resection (LAR) and circular double-stapled anastomosis without protective ileostoma. Since the pathomechanism of anastomosis leakage is still unclear, a suitable animal model would be most desirable.The objective was to assess the incidence of clinically apparent and inapparent leakage after LAR in pigs (n = 20). Endpoints were radiological, clinical, macroscopic, and histologic proof of anastomotic leakage on the 9th postoperative day. Integrity of anastomosis was assessed by double-contrast barium examination on 9th postoperative day. Animals were sacrificed and anastomoses were resected for histopathological investigation. In case of earlier clinical apparent anastomotic leakage, radiologic double-contrast barium was performed immediately.LAR with a circular double-stapled anastomosis without protective ileostoma was performed in 20 pigs (m:f = 8:12). Length of resection was 10-20 cm, anastomosis was performed 7 cm ab ano. Five animals (25%) developed clinical apparent anastomotic leakage (no appetite, fever, inactivity, tachypnea, discomfort, pain) between the 6th (n = 1) and 9th (n = 4) postoperative day, proven by double-contrast barium radiographs. Additionally in 1 animal clinical inapparent anastomotic insufficiency was observed radiologically. Total rate of leakage was 30% (n = 6). These results were confirmed by leucocytosis, low potassium levels, in two cases high ALT and AST and local peritonitis in all cases.Including one additional case of clinical inapparent leakage, total rate of anastomotic leakage was 30% (6/20). Thus we managed to establish a new experimental model of anastomotic leakage after low rectal resection comparable to the human situation.

The reported rate of clinically apparent anastomotic leakage (AL) in a low anterior resection of the rectum (LAR) (≤7 cm from the anal verge) using a circular double-stapled anastomosis (CDSA) without defunctioning stoma is up to 37.5 %. Since AL may result in life-threatening peritonitis, sepsis, and multiple organ failure, LAR and CDSA are regularly combined with defunctioning stoma. Accordingly, we now evaluated whether LAR and CDSA without defunctioning stoma but with extraluminal anastomotic application of an experimental fibrin sealant reduce the AL rate. This might prevent humans from defunctioning stoma increasing quality of life and decreasing surgical costs. Forty 8-week-old pigs underwent LAR and CDSA in an end-to-end technique (descendo-rectostomy). Animals were randomized into a therapy and control group (gr.). The therapy gr. (n = 20) received an additional extraluminal circular application of an experimental fibrin sealant to the anastomosis. The objective was to assess the incidence of clinically apparent and non-clinically apparent leakage through the ninth postoperative day. Double-contrast barium CT radiographs of the colorectal region were performed on the ninth postoperative day or earlier, in case there were clinical signs of AL. All remaining animals were sacrificed on the ninth postoperative day and the anastomotic region was histopathologically analyzed. In case of earlier diagnosed AL, animals were sacrificed immediately. Blood samples were taken for complete blood count, chemistry, and coagulation profile prior to surgery and on the first, third, fifth, seventh, and ninth postoperative day. A circular extraluminal anastomotic application of an experimental fibrin protection decreased the rate of clinically and non-clinically apparent AL from 20 % (n = 4) in the control group to 5 % (n = 1) in the treatment group. Ulcerations were also observed in both gr. (control gr.—5 animals, therapy gr. —3 animals). All animals with AL showed necrosis surrounding the hole at the anastomoses. Three additional animals had a full wall defect at the anastomotic region that was blocked by the experimental fibrin sealant. The fibrin sealant was present at necropsy in all treated animals. Circular anastomotic protection with the experimental fibrin sealant blocked anastomotic full wall defects, preventing peritonitis and significantly reducing the AL rate from 25 to 5 %.

The osteoconductive materials are important in bone regeneration procedures. Three dimensional (3D) reconstructions were obtained from the analysis. The aim of this study 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 two artificial matrixes inserted in previously artificially induced defects. For this study, under strict supervision 20 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 IngeniOss (for ten samples) and 4Bone(for the other ten samples). These materials were inserted into the induced defects. The femurs were investigated at 1 month, after the surgical procedures. The interfaces were examined using Time Domain (TD) Optical Coherence Tomography (OCT) combined with Confocal Microscopy (CM). 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 optical configuration uses two single mode directional couplers with a superluminiscent diode as the source centered at 1300 nm. 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 created due to the osteoinductive process. The TD-OCT has proven a valuable tool for the non-invasive evaluation of the matrix bone interfaces.

The problematic elements of bone regenerative materials are represented by their quality control methods. The defects repaired by bone grafting material were evaluated by en face optical coherence tomography and by synchrotron radiation micro-CT. The images obtained by efOCT show defects in some of the investigated samples, at the bone interface with different osteoconductive bone substitutes and we were able to detect gaps as small as 50 μm. After the common synchrotron radiation micro-CT investigations, the slides were reconstructed and the 3D model was obtained. Along with the possibility of navigating inside the structure, one big advantage of this technique was pointed out: the remaining regenerative materials can be separated from the normal bone and the new bone can be visualized. Optical coherence tomography can be performed in vivo and can provide a qualitative and quantitative evaluation of the bone augmentation procedure.

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.

Background Accumulating evidence shows the potential of bone marrow–derived endothelial colony-forming cells (bmECFCs) as promising tools for vascular repair. However, knowledge about their in vitro expansion, characterization, and functional behavior is still controversial. We demonstrate the in vitro generation of rat bmECFCs and analyze their ability to promote tissue reperfusion in a chronic hind-limb ischemia model. Methods Either in vitro–generated and characterized autologous bmECFCs or placebo was injected into ischemic hind limbs of Sprague-Dawley rats. Tissue perfusion was quantified by laser Doppler, in perfusion units (PU), at days 0, 15, and 30. Results Rat bmECFCs acquire a typical phenotype (CD34+VEGFR2+CD133+CXCR4+CD45−), culture, and functional behavior (Dil-ac-LDL+) in vitro. Injection of autologous bmECFCs improves tissue perfusion in ischemic hind limbs (183.5 ± 3.29 PUbmECFCs/day 30 versus 131 ± 3.9 PUcontrols/day 30, P Conclusions We conclude that rat bmECFCs promote ischemic tissue reperfusion and their proangiogenic properties are a potential mechanism for this effect.

Endoscopic internal mammary artery (IMA) approach represents a well-established procedure in coronary bypass surgery. However, such techniques require highly trained skills that need to be mastered in experimental models before taken into the clinic. To attain such skills, our aim was to develop a training model of thoracoscopic approach for harvesting of the IMA in pig. Ten pigs of either sex, weighing 20–28 kg, were used. The thoracic cavity was accessed with the animal in a lateral decubit position, through two ports of 10 mm and 1 of 5 mm, inserted on the anterior and posterior axillary lines immediately below the infrascapular angle into the pleural cavity, and a 30° Hopkins II telescope. The internal mammary artery (IMA) was identified and dissected from its origin in the subclavian artery until its distal part where it becomes the deep superior epigastric artery. Afterward, the arterial segment was clipped and harvested. The mean operative time was 73 min. Postoperative survival at 72 hours was 100% and 90% at 4 weeks. A mean length of 3.2 cm of IMA graft was obtained (2.8–4 cm). The pig IMA has no intercostal branches; thus, the mobilization of the vessel being greatly facilitated. The procedure has two critical steps: the proper trocar insertion and the dissection of the middle one-third of the IMA. Endoscopic-assisted IMA, harvesting in pigs, represents a useful tool for training in minimally invasive heart surgery and supports further development toward other clinical applications. © 2008 Wiley-Liss, Inc. Microsurgery, 2008.

Recent technical development has led to remarkable performances in video-guided surgical procedures. A video-endoscopic system (VES) is evaluated as an alternative magnifying solution for microsurgical procedures and compared to table-top microscopes in terms of technical and surgical aspects. Six surgical residents without microsurgical experience, alternating both systems, performed each 12 aortic end-to-end anastomoses on Sprague-Dawley rats using the triangulation technique. All anastomoses underwent quality review, total and single suture time, suture spacing, vessel bite, vessel overlapping and wall penetration were evaluated and graded. Overall anastomosis quality score was 52.28 (out of a maximum of 140) using the microscope and 42.7 using the VES. Despite significant differences in total anastomosis time, the learning curves are similar for the two systems and no major differences were noted in terms of overall anastomosis quality. Video-assisted microsurgery can become a useful instrument for microsurgery training. © 2007 Wiley-Liss, Inc. Microsurgery, 2007.

Plastic compounds have been used for several decades to generate anatomical constructs for the training of new surgeons and medical students alike. The present study seeks to highlight the advantages and disadvantages of two different plastic compounds (Technovit 7143 and Epoxi BIODURÒE12) used to create corrosion casts of the vascular branching patterns in free muscle flaps. Porcine latissimus dorsi muscle free flaps were used in this study to create corrosion casts of their vascular branching tree by injecting the two different plastic compound into the main arterial supply. The casts generated by Epoxy BIODURÒ E12 have superior qualities compared to the casts injected with Technovit 7143, because the injection process is smoother at all branching levels, without dilation, strictures or intramuscular extravasation of the injectable plastic compound. The corrosion casts resulted from injecting Epoxy BIODURÒ E12 exhibit better elasticity and better resistance to mechanical handling compared to the ones injected with Technovit 7143.

The intestinal mucosa is a major barrier in preventing bacteria invading the body but under certain circumstances mucosal gut barrier can be breached leading to the migration of bacteria to distant organs (mesenteric lymph nodes, spleen and liver). This process is termed bacterial translocation. The abnormal changes in gut ecosystem due to bowel resection led to bacterial overgrowth demonstrated postoperatively in male wistar rats duodenal and stool culture. We proved that after an extended bowel resection there is a direct relation between the number of the intestinal bacteria, the abnormal mucosal structural and functional changes and bacterial translocation to distant organs. A total of 22 male wistar rats divided in 4 groups were studied, short bowel syndrome was simulated in 16 by extended bowel resection (60%, 70% and 75%) while 6 rats had no surgery. Bacterial toxins induced local inflammation which altered neuromuscular tissue function, deteriorated further the gut barrier and increased bacterial overgrowth and bacterial translocation demonstrated by rat�s intestinal biopsy results. The sequence of events leading to translocation remained unclear. The correlation between the percentage of the duodenal aspirate Enterococcus and cecal stool bacteria in rats was R= 0.81, while the correlation between duodenal aspirate Enterococcus and cecal stool bacteria of rats without ileocecal valve resection compared to rats with ileocecal valve resection was R= 0.57 being statistically relevant. Simulated short bowel syndrome by extended intestinal resection caused bacteria overgrowth in residual rat�s bowel and this subsequently led to bacteria translocation to distant organs.

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.

Purpose: The aim of this study was to develop a method for standard laparoscopic access to the pudendal nerve in pigs to implant an electrode for chronic neuromodulation studies. Methods: Using routine laparoscopic surgical techniques, the pudendal nerve was located in 10 female pigs using standardized anatomical landmarks. A tined lead electrode was placed in parallel to the exposed pudendal nerve, and acute unilateral electrical stimulation was performed consecutively on both pudendal nerves. Bladder pressure and perineal skeletal muscle response was monitored during stimulation. Results: Standard access to the pudendal nerve was successfully established in the pig model with surgical times of approximately 45 minutes for bilateral electrode placement. Acute unilateral stimulation did not evoke bladder responses but resulted in reliable stimulation-dependent activity of the perineal skeletal muscles. The structural integrity of the pudendal nerves was confirmed in all cases. Conclusions: These results illustrate the effectiveness of laparoscopy for standardised, safe nerve localisation and electrode implantation at the pudendal nerve in pigs. Laparoscopic implantation represents an alternative approach for performing electrode implantation under optical guidance versus the standard approach of percutaneous, neuro-physiological monitored implantation. In the future, pudendal neuromodulation may be used as a supplement to sacral neuromodulation or as a standalone therapeutic approach, depending on the underlying bladder dysfunction.