Treatment of keloids and hypertrophic scars with cryotherapy

Keloid and hypertrophic scars are relatively frequent not wanted consequences of surgical treatment. 145 patients with unsatisfactory skin healing were treated in our department during last 5 years. When compared with corticotherapy and laser-therapy kryotherapy is very effective method. There were outstanding results with combination of cryotherapy and corticotherapy.     

Factor XIIIa-positive dermal dendritic cells in keloids and hypertrophic and mature scars

Keloids and hypertrophic scars have several common features. Both are reddish, firm, slightly protruding lesions that consist of proliferative fibroblastic cells and collagenous tissues. The aim of the present study was to evaluate the contribution of factor XIIIa (FXIIIa)-positive dermal dendritic cells to formation of keloids and hypertrophic scars. The numbers of FXIIIa-positive cells were counted in the keloid, hypertrophic scar and mature scar, each of which was divided into fibrocollagenous area and superficial dermal area overlying the nodular lesion. The features of the FXIIIa-positive cells were examined using immunohistochemical and immunoelectron microscopic techniques. More FXIIIa-positive cells were present in the overlying dermal area than in the fibrocollagenous area, commonly in three types of dermal lesion. The number of FXIIIa-positive dendritic cells was significantly greater in the overlying dermal area of keloids than in the corresponding area of hypertrophic or mature scar. Immunoelectron microscopic examination showed that the immunoreactivity for FXIIIa was seen mainly at the periphery of the cytoplasm of dermal dendritic cells. These results suggest that FXIIIa-positive dendritic cells in the overlying dermal area play active roles in formation of keloids and may contribute to dermal-epidermal interactions in keloids.     

肥厚性瘢痕和瘢痕瘤的病因病理学研究进展

在皮肤的愈合过程中，产生的瘢痕组织会高出皮肤表面，引起红、肿、热、痛及活动受限，甚者可导致严重的功能性问题或影响容貌，即所谓的瘢痕组织过度增生，包括肥厚性瘢痕和瘢痕瘤。本文对肥厚性瘢痕和瘢痕瘤的病因病理研究进行综述：     

A quantitative estimation of components in crayfish muscle fibres by stereological methods

The volume and surface densities of structural components in isolated long-sarcomere fibres of crayfish (Astacus fluviatilis) were estimated by stereological methods. The relative volumes were as follows (in % of the fibre volume): sarcolemmal invaginations (SI) 0.74; T-system 1.1; whole sarcoplasmic reticulum (SR) 4.7; junctional SR cisternae 0.41; mitochondria 1.1; nuclei 0.57; myofibrils 81.6; peripheral basal lamina 1.1; junctional gap 0.043; lysosomes 0.029. The values for surface densities were (in m²/100 m³): peripheral sarcolemma 1.27; SI 7.21; T-system 40.2; whole SR 146.4; junctional SR cisternae 14.88; junctional area of SR cisternae 7.05; mitochondria 8.98; nuclei 0.75; myofibrils 163. The invaginated membranes (SI and T-system) made up 97.4% of the total area of surface membranes and increased the effective area of peripheral sarcolemma 39 times. Using the values of the total electrical capacity from previous investigations onAstacus fibres, the specific capacity of surface membranes was calculated as 1.08 F/cm².     

Use of nude (athymic) mice for the study of hypertrophic scars and keloids: vascular continuity between mouse and implants

Hypertrophic scars and keloids appear to be unique to humans since animals are not known to form these lesions. Therefore, in an effort to develop an experimental model for their study, implants of these human lesions were made in nude (athymic) mice (nu/nu) in suprascapular subcutaneous pockets. The implants were recovered from 2 to 246 days. By histological and fine structural parameters all implants remained viable and their morphological character was maintained. Selected mice were injected with barium to confirm by microangiography vascular flow between mouse and implant. Hoechst stain for DNA, used to distinguish mouse cells from human cells, confirmed vascular anastamosis between host and implant: barium-filled vessels in the interior of the implant demonstrated human endothelial cells. Peripheral vascularization of the implant with minimal ingrowth of mouse vessels occurs during the first 8 days. Anastamosis probably occurs sometime before 16 days postimplantation, or earlier, depending upon the availability of patent microvessels in the implanted tissue. The presence of the implant does not appear to prompt a continuing vascular growth into or throughout the implant. The time frame of 16 days postimplantation should be taken into account when developing schemata of experimental or therapeutic modalities.     

Vascularization of the pineal complex in the lizard Tiliqua rugosa

The vascularization of the pineal complex in the lizard Tiliqua rugosa was investigated by vascular corrosion and latex casting techniques. The fine structure of the pineal capillaries was also studied by transmission electron microscopy. The pineal complex in T. rugosa consists of an elongated pineal gland proper and a separate, distinct parietal eye. The pineal complex derives an abundant blood supply from branches of the middle and posterior cerebral arteries. Scanning electron microscopy of vascular corrosion casts revealed a dense and extensive pineal capillary bed which drains ultimately into a wide longitudinal sinus suggesting an efficient pathway for the rapid removal of substances secreted by the gland. The parietal eye, which receives a unilateral left-sided blood supply from the unpaired anterior pineal artery, is shown to be a highly vascularized structure. The close morphological relationship between the pineal gland and dorsal sac, where the two structures apparently share the same blood vessels, suggests a functional relationship between them. The pineal capillaries are fenestrated with tight junctions between adjoining endothelial cells. Podia-like abluminal extensions of the endothelial cells were observed in close relation to unmyelinated nerve bundles. The basal margin of the pineal parenchyma is highly invaginated with thin finger-like cytoplasmic protrusions into the pericapillary space. Distinct bands of microfibrils form “struts” anchoring the pineal parenchyma to the endothelial wall. These features may have a role in the transfer of materials between the pineal gland and the blood stream. © 1993 Wiley-Liss, Inc.     

Perinatal asphyxia results in changes in presynaptic bouton number in striatum and cerebral cortex — a stereological and behavioral analysis

Deficits in cognitive function have been related to quantitative changes in synaptic population, particularly in the cerebral cortex. Here, we used an established model of perinatal asphyxia that induces morphological changes, i.e. neuron loss in the cerebral cortex and striatum, as well as behavioural deficits. We hypothesized that perinatal asphyxia may lead to a neurodegenerative process resulting in cognitive impairment and altered presynaptic bouton numbers in adult rats. We studied cognitive performance at 18 months and presynaptic bouton numbers at 22 months following perinatal asphyxia. Data of the spatial Morris water escape task did not reveal clear memory or learning deficits in aged asphyctic rats compared to aged control rats. However, a memory impairment in aged rats versus young rats was observed, which was more pronounced in asphyctic rats. We found an increase in presynaptic bouton density in the parietal cortex, whereas no changes were found in striatum and frontal cortex in asphyctic rats. An increase of striatal volume was observed in asphyctic rats, leading to an increase in presynaptic bouton numbers in this area. These findings stress the issue that volume measurements have to be taken into account when determining presynaptic bouton density. Furthermore, perinatal asphyxia led to region-specific changes in presynaptic bouton numbers and it worsened the age-related cognitive impairment. These results suggest that perinatal asphyxia induced neuronal loss, which is compensated for by an increase in presynaptic bouton numbers.     

Effects of combined renovascular hypertension and diabetes mellitus on myocardial cells,non-vascular interstitium and capillaries: a stereological study on rat hearts

Summary The effects of combined renovascular hypertension and diabetes mellitus on the rat heart were investigated in order to detect possible synergistic effects of the two conditions. Hypertensive diabetic and hypertensive non-diabetic animals were compared to diabetic and non-diabetic controls. Hypertension was established for 12 weeks by a surgical stenosis of the left renal artery; diabetes mellitus was maintained for 8 weeks by a single intraperitoneal injection of 60 mg/kg streptozotocin. Light microscopic stereology did not reveal significant divergences between diabetic hypertensives and non-diabetic hypertensives. Hypertension induced a focal perivascular and interstitial fibrosis with increased volume densities of non-vascular interstitium and fibrosis (P<0.001). Capillary density (Q_A) was decreased in transverse sections (P<0.01) and increased in longitudinal sections (P<0.01). This indicates a three-dimensional remodelling of the capillary bed with an increased number of obliquely running capillaries. At least the length density (L_V) of capillaries (mm/mm³) tends to be normalized in long-term renovascular hypertension. At the ultrastructural level, a synergism of hypertension and diabetes mellitus was observed: the volume ratio of mitochondria to myofibrils was significantly decreased in hypertensive diabetics, but not in non-diabetic hypertensives or in diabetics. This may enhance the risk of cardiac deterioration. We conclude that the primary target of the synergistic damage in hypertensive diabetic heart muscle disease is the myocardial cell and not the cardiac interstitium.Preliminary results of this study have been published in: Mall G (1991) Morphometric study on the rat heart in combined renovascular hypertension and diabetes mellitus. In: Nagano N, Dhalla NS (eds) The diabetic heart. Raven Press, New York, pp 115–124Dedicated to Prof. Dr. med. G. Seifert on the occasion of his 70th birthday     

Comparison of point counting and planimetry methods for the assessment of cerebellar volume in human using magnetic resonance imaging: a stereological study

The cerebellum is involved in motor learning and cognitive function in human. Many studies have been conducted to assess the cerebellar volume. To the best of our knowledge, there is no cerebellar volume study evaluating the efficiency and the accuracy of point-counting and planimetry methods of the Cavalieri principle in the literature. In this study, the volume of cerebellum was estimated in 53 Turkish young volunteers (26 males and 27 females), aged between 20 and 25 who are free of any neurological symptoms and signs, using serial magnetic resonance (MR) images. The cerebellar volumes of subjects were determined on MR images using the point-counting and planimetry methods. The mean results of planimetry method were 116.69 +/- 10.1 and 114.41 +/- 9.3 cm(3 )in males and females, respectively. The mean results of point-counting method were 116.34 +/- 10.6 and 113.48 +/- 8.8 cm(3 )in males and females, respectively. Our results revealed that female subjects had less cerebellar volumes compared with males, although there was no statistical significant difference between genders (P > 0.05). Total cerebellar volumes obtained by two different methods were not statistically different (P = 0.189) and they were correlated well to each other (r = 0.935). We found that the point-counting method takes less time than the planimetric method (mean 8 +/- 3.6 vs. 15 +/- 5.5 min). Thus, while planimetric and stereological approaches yield very similar results, the stereological method has the advantage of greater speed and, therefore, efficiency.     

A stereological approach to the study of neural organogenesis in Xenopus laevis

Summary Stereological methods are applied to the study of structural changes undergone, during neurulation, by the neuroepithelium of early embryos of Xenopus laevis. In light microscopy, we evaluate: the section area of the neuroepithelium, the nucleocytoplasmic ratio, the volume fraction of the intercellular spaces, of the nuclei and of the cytoplasm. In electron microscopy, the volumetric density and the surface ratio of mitochondria as well as the surface density of the endoplasmic reticulum are measured. Occasionally, the data found for Xenopus are compared to those found earlier in a similar study of the chicken embryo.     

Unbiased stereological quantification of unmyelinated fibers in the rat brain white matter

We applied unbiased stereological principles to estimate the volume of white matter and the total volume and length of the unmyelinated fibers in the white matter of the rat brain. The mean volume of the rat brain white matter was 119 mm³. The mean total length and the mean total volume of the unmyelinated fibers in the white matter were 323 km and 35 mm³, respectively. The described method provides a useful tool for future quantitative studies of changes in the unmyelinated fibers of white matter in various experimental conditions and in various neurodegenerative diseases.     

The proportionator: unbiased stereological estimation using biased automatic image analysis and non-uniform probability proportional to size sampling

The proportionator is a novel and radically different approach to sampling with microscopes based on the well-known statistical theory (probability proportional to size-PPS sampling). It uses automatic image analysis, with a large range of options, to assign to every field of view in the section a weight proportional to some characteristic of the structure under study. A typical and very simple example, examined here, is the amount of color characteristic for the structure, marked with a stain with known properties. The color may be specific or not. In the recorded list of weights in all fields, the desired number of fields is sampled automatically with probability proportional to the weight and presented to the expert observer. Using any known stereological probe and estimator, the correct count in these fields leads to a simple, unbiased estimate of the total amount of structure in the sections examined, which in turn leads to any of the known stereological estimates including size distributions and spatial distributions. The unbiasedness is not a function of the assumed relation between the weight and the structure, which is in practice always a biased relation from a stereological (integral geometric) point of view. The efficiency of the proportionator depends, however, directly on this relation to be positive. The sampling and estimation procedure is simulated in sections with characteristics and various kinds of noises in possibly realistic ranges. In all cases examined, the proportionator is 2-15-fold more efficient than the common systematic, uniformly random sampling. The simulations also indicate that the lack of a simple predictor of the coefficient of error (CE) due to field-to-field variation is a more severe problem for uniform sampling strategies than anticipated. Because of its entirely different sampling strategy, based on known but non-uniform sampling probabilities, the proportionator for the first time allows the real CE at the section level to be automatically estimated (not just predicted), unbiased-for all estimators and at no extra cost to the user.     

Keratinocyte-derived growth factors play a role in the formation of hypertrophic scars

In predisposed individuals, wound healing can lead to hypertrophic scar or keloid formation, characterized by an overabundant extracellular matrix. It has recently been shown that hypertrophic scars are accompanied by abnormal keratinocyte differentiation and proliferation, and significantly increased acanthosis, compared with normal scars. This study addressed the question of whether the development of normal and hypertrophic scars is regulated by differences in the growth factor profiles of both the epidermis and the dermis. The presence of interleukin-1alpha (IL-1alpha), IL-1beta, tumour necrosis factor-alpha (TNF-alpha), platelet-derived growth factor (PDGF), transforming growth factor-beta1 (TGF-beta1), and basic fibroblast growth factor (bFGF) was investigated in biopsies taken from breast reduction scars at 3 and 12 months following surgery. The samples were analysed by immunohistological methods and categorized as scars that remained hypertrophic (HH), became normal (HN) or remained normal after 12 months (NN). The epidermal expression of IL-1alpha was significantly increased in NN scars compared with HN and HH scars 3 and 12 months following operation, whereas the dermal expression showed no difference. PDGF was significantly increased in the dermis of normal scars after 3 months and in both the epidermis and the dermis of hypertrophic scars after 12 months. IL-1beta, TNF-alpha, TGF-beta and bFGF showed no differences. It is hypothesized that impaired production of keratinocyte-derived growth factors, such as IL-1alpha, leads to a decrease in the catabolism of the dermal matrix, whereas augmented epidermal PDGF production leads to increased formation of the dermal matrix in hypertrophic scars. These observations support the possibility that the epidermis is involved in preventing the formation of hypertrophic scars.     

A quantitative ultrastructural study of collagen fibrils in human skin normal scars,and hypertrophic scars

The diameter-distribution of collagen fibrils in the dermis of human skin and scar tissue has been investigated. In samples of normal skin, normal scar, and hypertrophic scar, prepared for transmission electron microscopy, collagen fibrils were systematically random-sampled and their diameters measured. There was a statistically significant difference between the diameter-distribution of fibrils in the papillary and reticular layers of dermis in normal skin. A similar pattern was found in normal scar, but not in hypertrophic scar. © 1995 WiIey-Liss, Inc.     

Age-dependent alterations in rat ventricular myocardium: a quantitative analysis

A quantitative electron microscopic examination of the ventricular myocardium of the male Fischer 344 rat was undertaken to determine the extent of age-dependent changes in ultrastructure. Rats of 6, 16 and 30 months of age were examined using a non-biased stereological test system. Volume fractions of mitochondria, myofibrillar mass and lipid remained unchanged during aging. Dense body volume fraction increased four-fold over the period studied, while the sarcoplasmic reticulum volume increased only from 6 months to 16 months and did not change thereafter. Biochemical analysis of the specific activities of the two lysosomal enzymes, acid phosphatase and beta-glucuronidase demonstrated a differential response to aging, with acid phosphatase remaining constant and beta-glucuronidase increasing slightly. Neither enzyme activity correlated with the pronounced change in dense body volume fraction. This study provides clear evidence that the previously observed changes in cardiac performance associated with aging are not readily explicable on the basis of a degradation of cardiac ultrastructure.