HISTOCHEMICAL AND MORPHOMETRIC ANALYSIS OF CONNECTIVE TISSUE IN HUMAN GLOMERULES DURING AGING

During aging there are alterations in human glomerules followed by increase of the connective tissue amount, accumulation of glomerular basal membrane and mesangial matrix on account of the rest of cell population. The aim of the study was quantification of the present connective tissue in mesangium of glomerules which morphologically did not show signs of sclerosis and determination of its part in glomerular structure. Investigation included 30 tissue samples of human kidneys of cadavers of both sexes, aged 20-85. Tissue samples were routinely stained with Mallory trichrome stain and analyzed by light microscope under 400´ magnifications. Images were analyzed with ImageJ software. Statistical analysis was performed with NCSS-PASS software. Cluster analysis was performed for the classification of glomeruli into 3 age groups, first with average age of 29, second with 44 and third with average of 71 years old. Histochemical investigations indicated the growth of connective tissue in mesangium of human glomerules during aging. It was also demonstrated morphometrically showing significant increase (p<0.05) of mean connective tissue area and its percentage in glomerule. In first, the youngest group the connective tissue was present in 17.33%, while in second group there was statistically significant increase (32.11%). The most significant growth compared to other age groups appeared in third group, where there was 40.66% of connective tissue in glomerule. Results of morphometrical and statistical analysis suggest that during aging process there is significant increase of area and percentage of connective tissue in the glomerule followed by reduction of cell siz

Morphometric Analysis of Nonsclerosed Glomeruli Size and Connective Tissue Content during the Aging Process

Number of sclerotic glomeruli increases during the aging process. Consequently, majority of remained nonsclerosed glomeruli become hypertrophic and some of them sclerotic, too. The aim of this study was to quantify the size and connective tissue content of nonsclerosed glomeruli and to evaluate the percentage of hypertrophic ones in examined human cases during the aging. Material was right kidney's tissue of 30 cadavers obtained during routine autopsies. Cadavers were without previously diagnosed kidney disease, diabetes, hypertension, or any other systemic disease. Tissue specimens were routinely prepared for histological and morphometric analysis. Images of the histological slices were analyzed and captured under 400x magnification with digital camera. Further they were morphometrically and statistically analyzed with ImageJ and NCSS-PASS software. Multiple and linear regression of obtained morphometric parameters showed significant increase of glomerular connective tissue area and percentage. Cluster analysis showed the presence of two types of glomeruli. Second type was characterized with significantly larger size, connective tissue content, and significantly lower cellularity, in relation to the first type. Such glomeruli might be considered as hypertrophic. First type of glomeruli was predominant in younger cases, while second type of glomeruli was predominant in cases older than 55 years

Morfološka, morfometrijska i imunohistohemijska analiza promena vezivnotkivnih omotača humanog suralnog nerva u toku starenja

Structural and functional age-related changes of peripheral nerve influence the regenerative capacity of the nerve. Stroma of the peripheral nerve acts as a support that maintains optimal microenvironment for the nerve impulse conduction, as well as the scaffold for the nerve fibers regeneration. Tissue samples of human sural nerve were analyzed using morphological, morphometric and immunohistochemical methods. The nerves were harvested during the autopsies and after lower limb amputation of the patients with diabetes mellitus and peripheral vascular disease. This study investigated the structural changes of the connective tissue sheaths during the ageing, as well as presence, localization and quantification of collagen IV, collagen I and laminin. Results of this study showed the presence of marked age-related changes in the perineurium and endoneurium of the investigated nerves, unlike epineurial sheath and fascicular structure of the nerve. Ageing is related to significant remodelling of the extracellular matrix of the perineurial sheath. Increased amount of collagen IV, as well as decreased expression of collagen I and laminin, were observed. Axonal atrophy in endoneurium were followed by more intense fibrosis and significant increase of collagen IV, as well as decreased amount of collagen I and laminin during ageing. In diabetic group we found significant endoneurial fibrosis with higher levels of collagen IV compared to the control, while patients with peripheral vascular disease suffered more prominent changes in the perineurial layer than age-matched control group, followed by increased deposition of collagen IV, collagen I and laminin. Observed differences in changes of the endoneurial environment and the perineurial sheath between senile, diabetic and vascular peripheral neuropathy further suggest the importance of extracellular matrix of nerve, and may contribute to better understanding of regeneration and functional recovery of nerve after injury, offering the guidelines to the future studies in the field of neurosciences and bioengineering

Morfološka, morfometrijska i imunohistohemijska analiza promena vezivnotkivnih omotača humanog suralnog nerva u toku starenja

Structural and functional age-related changes of peripheral nerve influence the regenerative capacity of the nerve. Stroma of the peripheral nerve acts as a support that maintains optimal microenvironment for the nerve impulse conduction, as well as the scaffold for the nerve fibers regeneration. Tissue samples of human sural nerve were analyzed using morphological, morphometric and immunohistochemical methods. The nerves were harvested during the autopsies and after lower limb amputation of the patients with diabetes mellitus and peripheral vascular disease. This study investigated the structural changes of the connective tissue sheaths during the ageing, as well as presence, localization and quantification of collagen IV, collagen I and laminin. Results of this study showed the presence of marked age-related changes in the perineurium and endoneurium of the investigated nerves, unlike epineurial sheath and fascicular structure of the nerve. Ageing is related to significant remodelling of the extracellular matrix of the perineurial sheath. Increased amount of collagen IV, as well as decreased expression of collagen I and laminin, were observed. Axonal atrophy in endoneurium were followed by more intense fibrosis and significant increase of collagen IV, as well as decreased amount of collagen I and laminin during ageing. In diabetic group we found significant endoneurial fibrosis with higher levels of collagen IV compared to the control, while patients with peripheral vascular disease suffered more prominent changes in the perineurial layer than age-matched control group, followed by increased deposition of collagen IV, collagen I and laminin. Observed differences in changes of the endoneurial environment and the perineurial sheath between senile, diabetic and vascular peripheral neuropathy further suggest the importance of extracellular matrix of nerve, and may contribute to better understanding of regeneration and functional recovery of nerve after injury, offering the guidelines to the future studies in the field of neurosciences and bioengineering

Morfološka, morfometrijska i imunohistohemijska analiza promena vezivnotkivnih omotača humanog suralnog nerva u toku starenja

Structural and functional age-related changes of peripheral nerve influence the regenerative capacity of the nerve. Stroma of the peripheral nerve acts as a support that maintains optimal microenvironment for the nerve impulse conduction, as well as the scaffold for the nerve fibers regeneration. Tissue samples of human sural nerve were analyzed using morphological, morphometric and immunohistochemical methods. The nerves were harvested during the autopsies and after lower limb amputation of the patients with diabetes mellitus and peripheral vascular disease. This study investigated the structural changes of the connective tissue sheaths during the ageing, as well as presence, localization and quantification of collagen IV, collagen I and laminin. Results of this study showed the presence of marked age-related changes in the perineurium and endoneurium of the investigated nerves, unlike epineurial sheath and fascicular structure of the nerve. Ageing is related to significant remodelling of the extracellular matrix of the perineurial sheath. Increased amount of collagen IV, as well as decreased expression of collagen I and laminin, were observed. Axonal atrophy in endoneurium were followed by more intense fibrosis and significant increase of collagen IV, as well as decreased amount of collagen I and laminin during ageing. In diabetic group we found significant endoneurial fibrosis with higher levels of collagen IV compared to the control, while patients with peripheral vascular disease suffered more prominent changes in the perineurial layer than age-matched control group, followed by increased deposition of collagen IV, collagen I and laminin. Observed differences in changes of the endoneurial environment and the perineurial sheath between senile, diabetic and vascular peripheral neuropathy further suggest the importance of extracellular matrix of nerve, and may contribute to better understanding of regeneration and functional recovery of nerve after injury, offering the guidelines to the future studies in the field of neurosciences and bioengineering

Morphometric Analysis of Connective Tissue Sheaths of Sural Nerve in Diabetic and Nondiabetic Patients

One of the most common complications of diabetes mellitus is diabetic neuropathy. It may be provoked by metabolic and/or vascular factors, and depending on duration of disease, various layers of nerve may be affected. Our aim was to investigate influence of diabetes on the epineurial, perineurial, and endoneurial connective tissue sheaths. The study included 15 samples of sural nerve divided into three groups: diabetic group, peripheral vascular disease group, and control group. After morphological analysis, morphometric parameters were determined for each case using ImageJ software. Compared to the control group, the diabetic cases had significantly higher perineurial index (P<0.05) and endoneurial connective tissue percentage (P<0.01). The diabetic group showed significantly higher epineurial area (P<0.01), as well as percentage of endoneurial connective tissue (P<0.01), in relation to the peripheral vascular disease group. It is obvious that hyperglycemia and ischemia present in diabetes lead to substantial changes in connective tissue sheaths of nerve, particularly in peri- and endoneurium. Perineurial thickening and significant endoneurial fibrosis may impair the balance of endoneurial homeostasis and regenerative ability of the nerve fibers. Future investigations should focus on studying the components of extracellular matrix of connective tissue sheaths in diabetic nerves

ABSOLUTE AND RELATIVE RENAL LENGTH IN CHRONIC KIDNEY DISEASES

The aim of the paper is to evaluate the significance of absolute and relative renal length in the diagnoses of several chronic kidney diseases (CKDs) in which kidney size changes in different manners during the disease course. The study included 181 patients: 35 with Balkan endemic nephropathy (BEN), 31 with diabetic nephropathy (DN), 30 with primary glomerular diseases (GN), 30 with autosomal recessive polycystic kidney disease (ADPKD), and 58 healthy controls (C). Absolute renal length was the distance between two most distant points on their poles and it was measured ultrasonographically, and relative length was obtained as the ratio of renal length and body height (kidney/body ratio, KBR). In the statistical analysis, One Way ANOVA test was used to establish the differences in absolute lengths and KBR between the studied groups; 2 test was used to establish the differences in the number of examinees of male and female gender; correlation and linear regression analysis were used to assess the association between age of the examinees and absolute and relative parameters of kidney size. The obtained results demonstrated that the average lengths of the right and left kidney were highest in ADPKD and lowest in BEN group. The average values of KBR of the right and left kidney showed a trend similar to that of average absolute lengths in all groups, except in GN and DN groups, in which absolute parameters of kidney size differed significantly from relative parameters. The correlation analysis showed that a significant negative correlation between age and absolute i.e. relative parameters of kidney size existed only in BEN group, but even in this case the differences between correlation coefficients of absolute and relative length of both kidneys were not statistically significant. Based on the obtained results, we could not establish the advantage of absolute over relative kidney length and vice versa in the studied CKDs. Further studies of larger patient samples with better gender and age distribution are therefore warranted

BILATERAL OSSIFICATION OF THE STYLOHYOID LIGAMENT

The stylohyoid ligament extends from the styloid process of the temporal bone to the lesser horn of the styloid bone. Various forms of ossification of this anatomical structure have been described in the literature. In our case, there was bilateral ossification of the stylohyoid ligament, a very rare occurrence. Ossification was partial on the right, and complete on the left side. The right stylohyoid ligament was only loosely attached to the styloid process, followed by a larger, ossified part of the attachment, finally attaching to the lesser horn of the hyoid bone in the form of a connective tissue bond. On the left, styloid process was present with its base and completely ossified stylohyoid ligament all the way to the lesser horn of the hyoid bone