Cell-Oriented Modeling of Angiogenesis

Due to its significant involvement in various physiological and pathological conditions, angiogenesis (the development of new blood vessels from an existing vasculature) represents an important area of the actual biological research and a field in which mathematical modeling proved particularly useful in supporting the experimental work. In this paper, we focus on a specific modeling strategy, known as “cell-centered” approach. This type of mathematical models work at a “mesoscopic scale,” assuming the cell as the natural level of abstraction for computational modeling of development. They treat cells phenomenologically, considering their essential behaviors to study how tissue structure and organization emerge from the collective dynamics of multiple cells. The main contributions of the cell-oriented approach to the study of the angiogenic process will be described. From one side, they have generated “basic science understanding” about the process of capillary assembly during development, growth, and pathology. On the other side, models were also developed supporting “applied biomedical research” for the purpose of identifying new therapeutic targets and clinically relevant approaches for either inhibiting or stimulating angiogenesis

Functional electrical stimulation of permanently denervated muscles, updated 2020

Spinal cord injury produces muscle wasting, which is especially severe after the complete and permanent damage of lower motor neurons that occurs in complete Cauda Equina Syndrome. Even in this worst-case scenario, we have shown that permanently denervated Quadriceps muscle can be rescued by surface Functional Electrical Stimulation and a purpose designed home-based rehabilitation regime. Here, our aim is to show that the effects are extended to both antagonist muscles and the skin of the thighs. Before and after 2 years of electrical stimulation, mass and structure of Quadriceps and Hamstrings muscles were quantitated by force measurements. Muscle gross cross section were evaluated using color computed tomography, muscle and skin biopsies by quantitative histology and immunohistochemistry. The treatment produced: a) an increase in cross-sectional area of stimulated muscles; b) an increase in muscle fiber mean diameter; c) improvements in ultrastructural organization; and d) increased force output during electrical stimulation. The recovery of Quadriceps muscle force was sufficient to allow 25% of the compliant subjects to perform stand-up and step-in place trainings. Improvements are extended to hamstring muscles and skin. Indeed, the cushioning effect provided by recovered tissues is a major clinical benefit. It is our hope that, with or without our advice, trials may start soon in Europe and Russia to provide persons-in-need the help they deserve

Lessons from Masters World Records: Lack of Gender Differences In Aging Muscle Decay Rates

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Dermal papillae flattening of thigh skin in Conus Cauda Syndrome

Our previous studies have shown that severely atrophic Quadriceps muscles of spinal cord injury (SCI) persons suffering with complete conus and cauda equina syndrome, and thus with permanent denervation-induced atrophy and degeneration of muscle, were almost completely rescued to normal size after two years of home based Functional Electrical Stimulation (hbFES). Since large surface electrodes were used to stimulate the denervated thigh muscles, we wanted to know if the skin was affected by this peculiar long-term treatment. Indeed, we demonstrated by two approaches that the epidermis decreases in thickness in the long term denervated persons, while it increased to almost pre-SCI values in hbFES compliant SCI persons. Here we report data of morphometry of skin biopsies from both legs of 18 SCI persons, harvested at enrolment in the Project RISE, to test if the Interdigitation Index, a simple measurement of the epidermal‐dermal junction, may provide a further precise quantitative evidence of the flattening of the skin in those SCI persons. The Interdigitation Index of the 36 skin biopsies shows a higly significant linear correlation with the years of SCI (p < 0.001). Furthermore, when the 18 SCI persons are divided in two groups (1 to 3.9 versus 4.1 to 8.0 years from SCI, respectively) and the data are compared, the later Group presents a statistically significant -22% decrease (p, 0.029) of the Interdigitation Index. On the other hand counting the papille do not provide the same strong evidence. In conclusion, the Interdigitation Index is an additional sound quantitative structural biomarker of skin atrophy and flattening occurring in SCI. The result correlates with the much severe extent of atrophy of the permanently denervated thigh muscles, as determined at both macro and microscopic levels.We are confident that the Interdigitation Index will provide sound evidence that the effects of hbFES, we previously reported on skeletal muscle and epidermis thickness, will be extended to the dermal layer of the skin, suggesting a coordinated negative effects of SCI on skeletal muscle and skin, and an improvement of both tissues after hbFES. Incoming analyses will be extended to basal lamina, collagene types, elastic fibers and skin annexes in the subcutaneous layer

Distribution of cannabinoid receptor 1 (CB1) and 2 (CB2) in subcutaneous tissue and fasciae

The endocannabinoid system is constituted by the endocannabinoid receptors (CB1 and CB2), by the endocannabinoids and the machinery for their biosynthesis and metabolism. Cannabinoid receptors have been localized in the central and peripheral nervous system as well as on cells of the immune system but recently they are discovered at epidermis and dermis1. The endocannabinoid system has been involved in different physiological processes, in particular many works in animal models have discovered an antinociceptive activities in inflammatory state and chronic inflammatory disease. Those findings suggest the possibility that the endocannabinoid system interacts with different cells so it will be interesting to provide a description of endocannabinoid receptors distribution. Immunohistochemical and molecular investigation for CB1 and CB2 localization was carried out in human skin and subcutaneous tissue and direct analysis on fibroblasts isolated from deep fascia2 and subcutaneous tissue of human and rat samples. The majority of endocannabinoid receptors were found in the keratinocytes of skin and mast cells close to subcutaneous adipose tissue, whilst in the deep fascia the presence is scarse. The CB2 receptors were more frequently highlighted respect to CB1 receptors. The abundant distribution of cannabinoid receptors on skin, mast cells and subcutaneous tissue provides implications for an anti-inflammatory, and this suggests more studies to evaluate the therapeutic potential of endocannabinoids

Muscle spindles of the rat sternomastoid muscle

The sternomastoid (SM) muscle in rodents presents a peculiar distribution of fiber types with a steep gradient from the ventral, superficial, white portion to the dorsal, deep, red region, where muscle spindles are restricted. Cross section of the medial longitudinal third of the rat SM contains around 10,000 muscle fibers with a mean diameter of 51.28±12.62 (μm +/- SD). Transverse sections stained by Succinate Dehydrogenase (SDH) reaction clearly presents two distinct regions: the dorsal deep red portion encompassing a 40% cross section area contains a high percentage of packed SDH-positive muscle fibers, and the ventral superficial region which contains mainly SDH-negative muscle fibers. Indeed, the ventral superficial region of the rat SM muscle contains mainly fast 2B muscle fibers. These acidic ATPase pH 4.3-negative and SDH-negative 2B muscle fibers are the largest of the SM muscle, while the acidic ATPase pH 4.3-positive and SDH-positive Type 1 muscle fibers are the smallest. Here we show that in thin transverse cryosections only 2 or 3 muscle spindle are observed in the central part of the dorsal deep red portion of the SM muscle. Azan Mallory stained sections allow at the same time to count the spindles and to evaluate aging fibrosis of the skeletal muscle tissue. Though restricted in the muscle red region, SM spindles are embedded in perimysium, whose changes may influence their reflex activity. Our findings confirm that any comparisons of changes in number and percentage of muscle spindles and muscle fibers of the rat SM muscle will require morphometry of the whole muscle cross-section. Muscle biopsies of SM muscle from large mammals will only provide partial data on the size of the different types of muscle fibers biased by sampling. Nonetheless, histology of muscle tissue continue to provide practical and low-cost quantitative data to follow-up translational studies in rodents and beyond

Exploring a tissue engineering strategy as a novel approach for haemophilic arthropathy treatment

Among the most disabling complications of Haemophilia, repeated and sponta- neous intra-articular haemorrhages may cause irreversible damage to the joint. This leads to haemophilic arthropathy, a polyarticular disease characterized by joint stiff- ness, chronic pain and a severely limited range of motion. Occurrence of haemophilic arthropathy can be avoided by the prophylactic administration of clotting factors to prevent articular haemorrhages, but it can also be addressed using anti-inflammatory drugs and surgery to alleviate the effects of articular damage, up to arthroplasty as resolute option [1]. However, innovative strategies for the prevention and treatment of this common and serious complication are still required, due to some important limits of current therapies, first of all inhibitor development. In this work, we inves- tigated a tissue engineering approach to regenerate articular focal lesions in Haemo- philic patients by in vitro development of an autologous bio-hybrid prosthesis. For this purpose, we isolated articular chondrocytes from Haemophilic patients (HaeCs) and characterized them for the first time in literature, to verify whether they were altered by blood exposure. Using healthy chondrocytes as control, optical microscope morphological analysis, flow cytometry immunophenotype evaluation and gene expression study by qRT-PCR were performed. After that, an innovative compos- ite scaffold was obtained by combining decellularized Wharton’s Jelly (W’s J) from human umbilical cord with a novel biodegradable polyvinyl alcohol (PVA) hydrogel [2]. Finally, we assessed HaeCs capacity to re-populate biosynthetic scaffolds by Scan- ning Electron Microscopy and MTT assay on cells seeded on supports. Taken togeth- er, our results contributed to define HaeCs phenotype, highlighting the possibility to use these cells for autologous implant. What is more, HaeCs capacity to growth and proliferate on composite scaffolds set the stage for planning the development of autologous tissue substitutes for haemophilic cartilage regeneration

L-citrulline is protective in hyperoxic lung damage and improves matrix remodelling and alveolarization

Moderate hyperoxia alters alveolar and vascular lung morphogenesis. Nitric oxide (NO) and matrix metalloproteinases (MMP) have a crucial role in the homeostasis of the matrix and bronchoalveolar structure and may be regulated abnormally by exposure to hyperoxia. Disruption of vascular endothelial growth factor (VEGF)-NO signaling impairs vascular growth and contributes to hyperoxia-induced vascular disease in bronchopulmonary dysplasia (BPD). We hypothesize that L-citrulline, by raising the serum levels of L-arginine and enhancing endogenous NO synthesis, might attenuate hyperoxia-induced lung injury in an experimental model of BPD. Neonatal rats (1 day old) were exposed to 60% oxygen or room air for 14 days and administered L-citrulline or a vehicle (sham). Lung morphometry were performed; Serum was tested for arginine level; Matrix metalloproteinases2 (MMP2) gene expression, VEGF gene and protein expression and endothelial NO synthase (eNOS) protein expression were compared. Mean linear intercept was higher in the hyperoxia and sham groups when compared with the room air (RA) and L-citr+hyperoxia treated group (p&lt;0.02). Secondary crests number was higher in L-citrulline treated and RA when compared to hyperoxia and sham group (p&lt;0.02). L-Arginine level rose in the L-citrulline-treated group (p&lt;0.05). L-citrulline did not affect MMP2 gene expression, but it regulated the MMP2 active protein, which rose in bronchoalveolar lavage fluid (p&lt;0.05), presumably due to a post-transductional effect. Compared with RA controls, hyperoxia significantly decreased VEGF and eNOS protein expression. At the same time, an increased lung VEGF gene and protein expression (p&lt;0.05) were also seen in the rats treated with L-citrulline. We conclude that: (i) hyperoxia decreases growth and disrupts VEGF-NO signaling of lung; (ii) the main effects of L-citrulline are an increased serum level of arginine, as a promoter and a substrate of the nitric oxide synthase; and (ii) a better alveolar growth and matrix control than in hyperoxia-induced lung damage seems promising