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Biomedical subjects

Alessandro Manelli

Publications and source records attributed to Alessandro Manelli.

9 recordsLinked to original sources

Whiplash injury and oculomotor dysfunctions: clinical-posturographic correlations.

Oculomotor dysfunctions are hidden causes of invalidity following whiplash injury. Many patients with whiplash injury grade II present oculomotor dysfunctions related to input disturbances of cervical or vestibular afferents. We used static posturography to investigate 40 consecutive patients with whiplash injury grade II and oculomotor dysfunctions. We demonstrated a relation between length and surface of body sway: the surface value (A) was higher than the length value (L) and this led to an open graph of body sway in the statokinesigram. Oculomotor rehabilitation can resolve the impairment of vestibular function but if therapy is delayed or the patient has been wearing an orthopaedic neck collar, more therapeutic sessions are required. In conclusion, without rehabilitation of the oculomotor muscles other therapies are not sufficient to recover the impairment caused by whiplash injury.

Adolescent↗

A new method for the joint visualization of vascular structures and connective tissues: corrosion casting and 1 N NaOH maceration.

Corrosion casting combined with scanning electron microscopy (SEM) has been widely used to study the morphofunctional aspects of microcirculation in many organs. In this study, we present an optimization of the corrosion casting (CC) technique associating it with NaOH 1 N maceration method to obtain a clear visualization of the relationships existing between the microvascular architecture of an organ and its extracellular matrix. Briefly, experiments were performed macerating the tissue previously injected with a low viscosity acrylic resin in 1 N NaOH and then observing it at SEM. In this study, we present an application of this technique to better evaluate the extracellular components of the vascular wall in medium-sized and capillary vessels both in skin and in kidney. The results obtained yielded clear images of the three-dimensional layout of medium-sized and capillary vessels in comparison with the extracellular environment. Furthermore, detailed information was obtained on the three-dimensional layout of fibers constituting the walls of venules, arterioles, and capillaries. In addition, the tubular collagenic structures surrounding the excretory tubules of the kidney and the dermal glands of the skin were depicted and their relationships with their vascular supply described in detail.

Animals↗

Tissue engineering techniques for the treatment of a complex knee injury.

We report and discuss the use of and rationale for tissue engineering techniques in a 40-year-old sportsman who suffered simultaneous anterior cruciate ligament (ACL) rupture, irreparable medial meniscal tear, and chondral lesion of the medial femoral condyle. A 2-step treatment was adopted to address all lesions. The first procedures consisted of ACL reconstruction and collagen meniscus implant (CMI), followed 6 months later by autologous chondrocyte implantation on a collagen membrane (MACI). A CMI biopsy was performed 6 months after implantation. Histologic and ultrastructural analysis documented scaffold invasion by cells and newly synthesized connective tissue. At 2-year follow up, the clinical and functional results were good and magnetic resonance imaging showed integration of the meniscal and cartilage implants. In this patient, each lesion needed to be addressed to achieve an optimal outcome. The primary goal was to restore normal joint biomechanics by performing CMI and ACL reconstruction. MACI was an adequate solution for the management of the large (5 cm(2)) chondral defect.

Adult↗

The 3D structure of crimps in the rat Achilles tendon.

The ultrastructure of crimps of the Achilles tendon of rat, excised and processed in a slack condition, was investigated by atomic force microscopy in air, in fluid and by scanning electron microscopy and stereo reconstruction. The tendon was made of distinct fascicles, each comprising a succession of straight segments connected by sharp angles. The length of the segments and the interposed angles varied widely. In particular, the angles ranged from almost zero to over 135 degrees . We did not observe a unique structure for the hinge regions, but rather a variety of gradations of buckling and/or torsion with no evident correlation with other features of tendon. A constant hallmark was the local loss of regular molecular packing, as revealed by the disappearance of the D-banding. Our results do not support recent reports of a helical structure or smooth sinusoidal waves in tendons. Such structures may nonetheless exist in other non-tensile structures whose collagen fibrils exhibit a helical inner architecture and are able to follow a highly convoluted course without buckling or crimping.

Achilles Tendon↗

The extracellular matrix of the human aortic wall: ultrastructural observations by FEG-SEM and by tapping-mode AFM.

Fragments of human ascending aorta harvested during heart surgery were cryofractured and observed by scanning electron microscopy (SEM) and atomic force microscopy (AFM). Elastic fibers appear as irregular, undulated laminae of variable size and shape. Their surface shows an evident fibrous texture suggestive of a criss-crossed, delicate filamentous scaffold and is marked by a number of features such as ridges, holes and protruding ribs. At higher magnification, both SEM and AFM show the surface composed of a finely granular material, with a bead size of approximately 20 nm. However, the thickness of the metal coating in one case, and the tip convolution effect on the other, may equally result in an artifactual enlargement of the structures, so that the beads may be significantly smaller. The surfaces created by the fracture always appear smooth and compact and with this technique do not reveal significant detail. The collagen component is mostly represented by small, uniform fibrils gathered in flexuous bundles and following a wavy course not unlike that of the elastic laminae. An orthogonal lattice of small proteoglycans is readily evident even without a specific treatment. Occasionally, the fibrils appear encrusted or engulfed in a grainy matrix reminiscent of the elastic fiber surface. Fluid Tapping-Mode Atomic Force Microscopy simultaneously reveals the surface-bound proteoglycans and the inner architecture of the fibrils, composed of smaller subunits following a spiral course with a winding angle of approximately 17 degrees.

Aged↗

Histology and ultrastructure of a tissue-engineered collagen meniscus before and after implantation.

The collagen meniscus implant (CMI) is a tissue-engineering technique designed to stimulate regeneration of meniscus-like tissue in cases of irreparable tears or previous meniscectomy. CMI morphology was investigated before and after implantation by light microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). In a case series biopsy specimens were harvested from four patients who underwent a second arthroscopic look 6 months after placement of the CMI. CMI sections appeared composed of parallel connective laminae of 10-30 microm, connected by smaller bundles (5-10 microm). This connective network formed lacunae with diameters between 40 and 60 microm. At greater magnification, the walls of the lacunae demonstrated tightly packed and randomly distributed collagen fibrils, with diameters ranging from 73 to 439 nm. In the biopsy specimens, the lacunae were filled with connective tissue that contained newly formed vessels and fibroblast-like cells, presenting an abundant rough endoplasmic reticulum and several mitochondria. In the extracellular matrix, the collagen fibrils showed uniform diameters (126 nm +/- 32 nm). The original structure of CMI was still recognizable, and no inflammatory cells were detected within the implant. The morphological findings of this case series demonstrate that CMI provides a three-dimensional scaffold suitable for colonization by precursor cells and vessels and leading to the formation of a fully functional tissue.

Adult↗

A vision-based, 3D reconstruction technique for scanning electron microscopy: direct comparison with atomic force microscopy.

High-resolution, detailed 3D reconstructions of biological specimens obtained from scanning electron microscopy stereo-micrographs and proprietary software were compared with Tapping-Mode AFM datasets of the same fields. The reconstruction software implements several original solutions including a neural adaptive point-matching technique, the ability to build an irregular triangulated mesh rather than a regular orthogonal grid, and the ability to re-map one of the original images exactly onto the reconstructed surface. The technique was applied to human nerve tissue to obtain 1,424 x 968-pixel, texture-mapped datasets, which were subsequently compared against 512 x 512-pixel AFM datasets from the same viewfields. Accounting for the inherent differences of the two techniques, direct comparison revealed an excellent visual match. The correspondence was also quantified by calculating the cross-correlation coefficient between corresponding altimetric profiles in SEM and AFM data, which consistently exceeded a figure of 0.9, with a rate of point mismatch in the order of 0.01%. Research is still underway to improve the robustness of the technique when applied to arbitrary images

Humans↗

The collagenic structure of human digital skin seen by scanning electron microscopy after Ohtani maceration technique.

We performed a morphological scanning electron microscope (SEM) study to describe the fine structure and disposition of collagenous tissue in the human toe. After therapeutic amputation of a human right leg, we applied the Othani maceration technique to the skin of three toes surgically explanted from the foot. We distinguished eight cutaneous regions and focused on some specialized collagenous structures differing in the thickness of the skin. The eight areas investigated were: the dorsal skin, the eponychium, the perionychium, the hyponychium, the region under the visible nail, the nail root, the plantar skin and finally the toe tip. Each of these areas is characterized by a distinctive collagenous surface disposition, with some peculiar features mostly related to dermal papillae. At high magnification, we observed the spatial arrangement of the collagen fibers constituting the top of the dermal papillae that represents the attachment site of the proliferative basal layer of the epidermis. We also noted an impressive density of collagen fibers throughout the thickness of the dermal layer, organized in specialized structures and constituting the skeleton of dermal thermoreceptorial corpuscles or sweat glands. A combination of SEM and Ohtani technique disclosed the three-dimensional architecture of the collagenous matrix of tarsal skin under physiologic conditions, giving a detailed description of the most reactive tissue during pathologic processes.

Amputation, Surgical↗

Structure and ultrastructure of microvessels in the kidney seen by the corrosion casting method.

Scanning electron microscopic observation of corrosion casts is the finest technique to describe spatial patterns of microvessels in many organs, giving a readily interpreted representation of their vascular architecture without interference from surrounding tissues. We focused on the renal cortex of guinea pigs to make an in-depth morphological analysis of structural and ultrastructural details left by the cells on the resin cast. In addition, we made a qualitative description of normal variants usually observed in glomerular disposition, arteriolar morphology or capillary arrangement in the space to shed more light on the relationship between vascular tissue and surrounding cells. The study also disclosed some examples of vascular adaption to physiological and pathological conditions occurring in renal microvessels such as many systems essential to flow regulation, filtration and excretory processes. At lower magnification, all major vessels can be readily distinguished: interlobar, arciform and interlobular arteries and veins, along with a web of peritubular and capsular capillaries. At higher magnification, the glomeruli become visible and the afferent and efferent arteries and the tortuosity the inner vessels can be distinguished. In some of them, the resin, due to the narrowing sizes, suddenly stopped leaving a half-casted glomerulus. This helped to reveal its internal circulation characterized by thin capillaries with a high degree of bi or trifurcation. In addition, we confirmed the close correspondence between cellular ultrastructural detail (pores, corrugations of cellular membrane, perivascular cell branches) and the impressions left on the resin visible only at high magnifications.

Adaptation, Physiological↗