In vitro evidences of different fibroblast morpho-functional responses to red, near-infrared and violet-blue photobiomodulation: Clues for addressingwound healing

Although photobiomodulation (PBM) has proven promising to treat wounds, the lack of univocal guidelines and of a thorough understanding of light–tissue interactions hampers its mainstream adoption for wound healing promotion. This study compared murine and human fibroblast responses to PBM by red (635 ± 5 nm), near-infrared (NIR, 808 ± 1 nm), and violet-blue (405 ± 5 nm) light (0.4 J/cm2 energy density, 13 mW/cm2 power density). Cell viability was not altered by PBM treatments. Light and confocal laser scanning microscopy and biochemical analyses showed, in red PBM irradiated cells: F-actin assembly reduction, up-regulated expression of Ki67 proliferation marker and of vinculin in focal adhesions, type-1 collagen down-regulation, matrix metalloproteinase-2 and metalloproteinase-9 expression/functionality increase concomitant to their inhibitors (TIMP-1 and TIMP-2) decrease. Violet-blue and even more NIR PBM stimulated collagen expression/deposition and, likely, cell differentiation towards (proto)myofibroblast phenotype. Indeed, these cells exhibited a higher polygonal surface area, stress fiber-like structures, increased vinculin- and phospho-focal adhesion kinase-rich clusters and α-smooth muscle actin. This study may provide the experimental groundwork to support red, NIR, and violet-blue PBM as potential options to promote proliferative and matrix remodeling/maturation phases of wound healing, targeting fibroblasts, and to suggest the use of combined PBM treatments in the wound management setting

Intraductal papillary mucinous neoplasia (IPMN) of the pancreas: the pivotal role of MRI for the differential diagnosis and the choice of treatment

Macrocystic pancreatic tumors seem to play an important role among neoplastic lesions of the pancreas as they sometimes either show a malignant potential or they already have neoplastic foci inside the cystic tumor. Differential diagnosis is a key factor in comparison with other cystic tumors which are not malignant as Serous Cystic Tumors (SCTs) and Mucinous Cystic Tumors (MCTs). So diagnostic imaging has become more and more important. Since May 2009 we have observed more than 200 patients with cystic lesions of the pancreas. All the patients underwent a CholangioPancreato MagneticResonance (CPMR) after an Ultrasound and/or a CT scan. Then we excluded from our study solid lesions, pseudocysts and tumors with clear signs of malignancy. CPMR was sometimes performed also using a secretine test. Finally 51 patients were evaluated and underwent a follow up programme till now. Among these patients we found 34 Intraductal Papillary Mucinous Neoplasia (IPMN), 7 MCTs and 10 SCTs. As we know that all SCTs show a lobulated septate pattern, differential diagnosis with IPMN is mandatory in order to give to the patient the treatment of choice. CPMR revealed in 32 out of 34 IPMN patients a communication between the lesion and the main pancreatic duct (MPD); so this sign, which is patognomonic of IPMN neoplasia, confirmed the diagnosis. All lesions > than 3 cm were resected by surgery (4 MCTs and 3 IPMN). Definitive histology always confirmed preoperative diagnostic imaging. Now the patients are all disease free at follow up. The other 44 patients undergo CPMR every 6 months following a “wait and see” policy. CPMR seems to be fundamental for the diagnostic screening of IPMN. This is a simple, safe and non invasive procedure which allows an early diagnosis and a better chance of cure for this kind of patients

Structural organization of enteric

The organization of the Enteric Nervous System (ENS) was studied in the human colon. Fragments of the whole colonic wall were either routinely processed or Zinc-Iodide Osmium impregnated. Single-layer preparations were also obtained from some of the Zinc-Iodide Osmium-impregnated specimens. The results showed some differences in the organization of human colonic ENS from that of other mammals. In fact, the human submucous plexus was made up of three interconnected ganglionated networks arranged along three different planes. With respect to the myenteric plexus, its ganglia were large sized and irregularly shaped. Moreover, during the microdissection of the colonic wall, we found the absence of a cleavage plane between the circular and longitudinal muscle layers; on the other hand the cleavage plane between mucosa and submucosa was not immediately below the muscularis mucosae, but slightly deeper, since the innermost part of the submucosa remained adhering to overlying layers

Cytocompatibility of polyurethane foams as biointegrable matrices for the preparation of scaffolds for bone reconstruction

This work reports preliminary results on the development of biointegrable scaffolds, composed of biostable 3D polymer matrices and bioabsorbable inorganic salts, to be used for cell anchorage in bone regeneration. Three crosslinked polyurethane foams (PUFs), prepared by one-step bulk polymerisation from a polyether-polyol mixture, polymeric MDI and water as expanding agent, were tested for their ability to promote adhesion and growth of bone-derived cells. The open porosity of these foams ranged from 16 to 31% with an average pore size of 470÷600 μm, compressive strength (at 10% ε) of 0.28÷0.38 MPa and elastic moduli of 4.88÷6.61 MPa. The human osteosarcoma line Saos-2, and primary cultures of normal human articular chondrocytes and bone marrow-derived (HBM) stromal cells were used for in vitro cytocompatibility tests. For cell adhesion and proliferation analysis, DNA synthesis was evaluated by 3H-thymidine uptake. Osteoblastic differentiation of Saos-2 adherent cells was determined by measuring the enzymatic activity of alkaline phosphatase (ALP). All cell types were able to adhere to all tested PUFs and to synthesize DNA. At 48 hr culture, HBM stromal cells showed the maximal rate of adhesion with the highest rate of proliferation onto PUFs with the largest pore size, whereas both chondrocytes and Saos-2 appeared to adhere preferentially onto foams exhibiting the highest percentage of open porosity. Up to 8 days in culture Saos-2 cells were able to proliferate into all PUFs, with a time-dependent increase of DNA synthesis and ALP activity. At SEM, the morphology of cells adherent to PUF pores was spread with cytoplasmatic extroflessions, indicating a good metabolic activation. These results demonstrate a good cytocompatibility of the proposed 3D matrices, suggesting that their use in the preparation of composite scaffolds is worth further investigation. (Journal of Applied Biomaterials & Biomechanics 2003; 1: 58-66

Vitamin E prevents neutrophil accumulation and attenuates tissue damage in ischemic-reperfused human skeletal muscle

Neutrophil accumulation and the consequent production of oxygen-derived free radicals are involved in the pathogenesis of Ischemia-Reperfusion syndrome. In this study we investigated whether a treatment with Vitamin E, which has antioxidant properties, could attenuate the tissue damage by interfering with the influx of neutrophils within the ischemic and reperfused human skeletal muscle. To this purpose, patients undergoing aortic crossclamping during the surgical repair of aortic abdominal aneurysm were studied as a model of ischemiareperfusion of the lower limb muscles. Muscle biopsies from the right femoral quadriceps of patients not receiving and receiving Vitamin E pretreatment before surgery were taken: a) after the induction of anaesthesia, as control samples, and b) after a period of ischemia followed by 30 min of reperfusion. The tissue samples were either routinely processed for morphological study and immunohistochemical analysis to detect an altered expression of specific endothelial adhesion proteins, such as E-selectin and ICAM-1. The results obtained showed that Vitamin E administration was able to prevent the accumulation of neutrophils within the ischemic and reperfused muscle. This beneficia1 effect of Vitamin E was due to its ability to hinder the expression of E-selectin and ICAM-1, molecules known to increase the adhesiveness of endothelium to circulating neutrophils. After treatment with Vitamin E a marked attenuation of the reperfusion injury was also evident. In conclusion, Vitamin E treatment may be considered a valuable tool for protection against the ischemiareperfusion damage of human skeletal muscle