Biomedical subjects

A Dziedzic-Goclawska

Publications and source records attributed to A Dziedzic-Goclawska.

At least 19 recordsLinked to original sources

PubMed1999

Amnion allografts prepared in the Central Tissue Bank in Warsaw.

Applications of allogenic amnion grafts range from wound dressing of severe burns, dermabrasions and lower extremity ulcer treatments to plastic surgery, laryngology and ophthalmology. The aim of the present study was to elaborate the method of processing, preservation and sterilization of human amnion allografts prepared as wound dressing used mainly for burned patients. During the amniotic sac processing (after separation of chorion) special attention was paid to ensure that the epithelial side of amnion is placed directly on polyester net used as a support. After application on the wound, the epithelial side with the basement membrane is facing outwards; this will promote migration, attachment and spreading of the host cells encouraging epithelialization. Human amnion allografts were preserved by lyophilization or deep-freezing and subsequently radiation-sterilized with a dose of 35 kGy. It has been observed, however, that lyophilized irradiated allografts are resorbed within a few days, while frozen irradiated ones better adhere to wound and persist even 3 weeks after grafting, therefore, it has been decided to preserve amnion by deep-freezing. Since the beginning of 1998 over 400 preserved radiation-sterilized amnion allografts (with a total surface area over 40,000 cm2) have been prepared at the Central Tissue Bank in Warsaw and distributed to clinics and hospitals throughout the country.

Amnion↗

PubMed1997

Alteration of mineral crystallinity and collagen cross-linking of bones in osteopetrotic toothless (tl/tl) rats and their improvement after treatment with colony stimulating factor-1.

A common feature of various types of mammalian osteopetroses is a marked increase in bone mass accompanied by spontaneous bone fractures. The toothless (tl/tl) rat osteopetrotic mutation is characterized by drastically reduced bone resorption due to a profound deficiency of osteoclasts and their precursors. An altered bone morphology has also been observed. The mutants cannot be cured by bone marrow transplantation, but skeletal defects are greatly reduced after treatment with colony stimulating factor 1 (CSF-1). The objectives of this study were to characterize mineral and collagen matrices in cancellous and compact bone isolated from long bones of 6-week-old normal littermates, tl/tl osteopetrotic mutants and mutants (tl/tl) treated with CSF-1. There were no differences in bone mineral content, but a significant decrease in the crystallinity of mineral evaluated by the method based on electron paramagnetic resonance spectrometry was observed in all bones of tl/tl mutants as compared to that of controls. Within the collagen matrix, slight decreases in the labile cross-links, but significant increases in the content of the stable cross-links, pyridinoline, and deoxypyridinoline, were observed in both cancellous and compact bone of osteopetrotic mutants. In tl/tl mutants treated with human recombinant CSF-1, the normalization of the crystallinity of bone mineral as well as collagen cross-links was found. Our results indicate that remodeling of bone matrix in tl/tl mutants is highly suppressed, but that after treatment with CSF-1, this activity recovers significantly. Taken together, these data provide further support for the hypothesis that CSF-1 is an essential factor for normal osteoclast differentiation and bone remodelling.

Animals↗

PubMed1996

Optical diffractometry in bone research.

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Bone and Bones↗

PubMed1994

Bone powder exposed to the action of 12C and 25Mg ion beams as investigated by electron paramagnetic resonance spectroscopy.

Bovine compact bone powder was irradiated with alpha-particles of 241Am, 12C6+ and 35Mg12+ ion beams, then examined by the e.p.r. spectroscopy. The stable e.p.r. signal found is of the same characteristic as that observed in gamma-irradiated bone (g perpendicular = 2.0017, g parallel = 1.9973, delta Hpp = 0.85 mT). Thus, the structure of radiation induced paramagnetic center in bone does not depend on the LET of radiation used. The radiolytic yields of the formation of stable paramagnetic centres in bone powder irradiated with gamma-rays are G gamma = 0.08 +/- 0.01 while with 12C6+ ion beams roughly about Gc = 0.02, respectively.

Animals↗

PubMed1992

Decreased heterotopic osteogenesis in vitamin-D-deficient, but normocalcemic guinea pigs.

The effect of vitamin D deficiency unhampered by hypocalcemia on de novo bone formation was studied in guinea pigs. Heterotopic induction of osteogenesis was evaluated 4 weeks after intramuscular transplantation of allogenic urinary bladder transitional epithelium from vitamin-D-repleted (+D) donors into +D and -D recipients. In -D recipients the frequency of osteogenesis and the amount of induced bone were significantly diminished; induced bone was less mature, scantly cellular woven bone poorly repopulated with bone marrow. No effect of vitamin D deficiency on orthotopic bone growth and on mineralization of orthotopic and heterotopically induced bone was observed. It is proposed that in addition to inducing factors (BMPs, growth factors) which may be responsible for transformation of mesenchymal cells to osteoprogenitor cells, normal concentrations of 1,25-(OH)2D3 may be required for proliferation and further differentiation of these cells into osteoblasts and for expression of genes engaged in extracellular matrix formation and maturation.

Alkaline Phosphatase↗

PubMed1991

Radiation-induced paramagnetic centers in research on bone physiopathology.

It has been found that ionizing radiation induces stable paramagnetic centers in the crystalline lattice of bone hydroxyapatite (HA). These defects, which can be detected and measured by electron spin resonance (ESR) spectrometry, can be treated as a new kind of biologic marker and used for the following: (1) evaluation of crystallinity of bone mineral, because the yield of these centers depends on the size of HA crystals; (2) quantification of the rebuilding process of radiation-sterilized bone grafts, because of the stability of the centers; and (3) estimation of the dose of ionizing radiation absorbed by living organisms, because of the linear relation between the dose of ionizing radiation and the yield of radiation-induced paramagnetic centers in bone mineral.

Animals↗

PubMed1991

Effect of radiation sterilization on the osteoinductive properties and the rate of remodeling of bone implants preserved by lyophilization and deep-freezing.

The effect of various doses of ionizing radiation on the osteoinductive properties of decalcified bone matrices implanted heterotopically and on the rate of remodeling of nondecalcified bone grafts implanted orthotopically in allogeneic systems was studied. Decalcified bone matrices and nondecalcified bone grafts were preserved by lyophilization or by deep-freezing and were subsequently irradiated with appropriate doses at room temperature or at -72 degrees. Lyophilized matrices irradiated at room temperature with 35 and 50 kGy, respectively, were completely resorbed five weeks after heterotopic implantation into the muscles and did not induce osteogenesis, whereas the resorption of deep-frozen ones irradiated with the same doses at -72 degrees was slower and new bone formation was induced. The preservation of the osteoinductive capacity of irradiated, deep-frozen matrices may depend on two factors: reduction of radiation damage on the inducing agents and collagen irradiated in the presence of water, which may diminish the rate of matrix resorption. The rate of remodeling of undecalcified deep-frozen bone implants irradiated at -72 degrees and grafted orthotopically was higher than that of lyophilized ones irradiated at room temperature. It is possible that the temperature during irradiation plays a critical role in protection against radiation damage.

Animals↗

PubMed1989

Spreading of cells on various substrates evaluated by Fourier analysis of shape.

In order to evaluate in mathematical terms the morphological changes occurring in the course of cell spreading, Fourier analysis of shape was applied. Human urothelial Hu 961 b cells plated on type IV collagen, fibronectin, laminin, glass and bovine serum albumin (BSA) were studied. Fourier parameters describing cell shape as well as surface areas covered by the cells on the substrate were subjected to statistical analysis. Using analysis of variance and discriminant analysis it was found that parameters describing cell shape (both gross shape of cells and their fine scale contour foldings) possessed a higher power of discrimination between the cells spread on various substrates than the differences in cell surface areas. In the course of observation (75 and 150 min) the highest number of attached cells and highest degree of spreading were found when cells were plated on type IV collagen. Moderate alterations in cell shape and moderate increase of surface area were seen in the group of cells seeded on fibronectin, whereas the cells plated on laminin, glass and BSA revealed a moderate increase of surface area, but no changes in their shape were observed. The differences in attachment of cells and in the degree of their spreading might be due to the variation in expression of plasma membrane receptors for various substrates. The Fourier analysis of cell shape coupled with measurement of surface area is a good tool for quantitative evaluation of cell spreading and can be used for discrimination between cells spread on different substrates.

Cell Adhesion↗

PubMed1988

Differences in the kinetics of the mineralization process in endochondral and intramembranous osteogenesis in human fetal development.

About 300 samples of bone tissue from femoral diaphyses and from parietal bones of calvaria were collected from human fetuses aged from 6-8 to 40-41 weeks in order to compare the kinetics of the mineralization process in endochondral and intramembranous bone formation. The crystallinity of bone mineral was evaluated by the method based on electron spin resonance (ESR) spectrometry, and mineral content was measured after ashing of the samples. Highly significant differences in the kinetics of deposition of mineral (ash content) and maturation of deposited mineral (crystallinity coefficient) were found between parietal bones and femoral diaphyses in the course of fetal development. In the early stages of development (6-8 to 10-12 weeks), the content of mineral and its crystallinity in parietal bones were significantly lower than in femoral diaphyses. In the parietal bones a gradual increase in both values was observed until they reached a plateau at the age of 30 weeks. On the other hand, in femoral diaphyses of 6-8-week-old fetuses mineral content was high and deposited mineral showed a high degree of crystallinity. Both these values did not change significantly until birth. The differences in kinetics of the mineralization process between the weight-bearing bones and bones that are not exposed to the influence of strong external forces are most probably genetically conditioned. The collected data supply embryological information and might be used in the future for evaluation of pathologic changes in skeletal development.

Embryonic and Fetal Development↗

PubMed1988

Effect of 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) and dichloromethylidene-bisphosphonate (Cl2MBP) on the structure of the organic matrix of heterotopically induced bone tissue.

The effect of 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) and dichloromethylidene-bisphosphonate (Cl2MBP) on the structure of the organic matrix of heterotopically induced bone in guinea pig was studied. Heterotopic bone formation was induced by transplantation of allogenic urinary bladder epithelium. Starting from the day of transplantation the animals were treated subcutaneously with HEBP and Cl2MBP with a dose of 12.5 mg P/kg/day during 35 days. The control group was injected with 0.9% NaCl solution. The advantage of heterotopic bone induction as an experimental model is the fact that the applied drugs act on de novo bone formation. Collagen fibers were treated as markers of bone because their size and spatial arrangement reflect the structure and maturity of organic matrix of this tissue. Decalcified histological sections of induced bone, taken 35 days after implantation of inductor, were stained by the picrosirius method. This staining enhances the natural birefringency of collagen fibers and allows for better and specific visualization of collagen fibers bundles under polarizing microscope. In this way the amount of information in the analysed image is increased. Thirty five microphotographs were analysed from each of the investigated groups with the use of optical diffractometry. The radial distribution of light intensity in diffraction patterns was analysed what allowed to evaluate spatial frequencies connected with the width of collagen bundles in induced bone tissue. Since the spatial arrangement of collagen fibers in newly formed bone is random, analysis of angular distribution of light intensity in diffractograms was not performed. Using discriminant analysis the significant differences between all three studied groups of animals were found.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1986

Numerical evaluation of changes in the cytoplasmic microtubule complex of C3H mouse cells by optical diffractometry and of changes in cell shape by Fourier analysis.

MO mouse cells in culture on glass were treated with taxol, or nocodazole, or incubated at 4 degrees C to alter their cytoplasmic microtubule complex (CMTC). From each treated group and from an untreated group, 30 cells stained with an antiserum against tubulin, were photographed under the photomicroscope, and negatives were analysed by optical diffractometry. Differences between groups of cells were tested by variance analysis. Phase-contrast micrographs of the same cells were used for Fourier analysis of cell shape. Both types of analyses provided numerical objective data about changes in the CMTC and in cell shape that were typical for the kind of treatment. We conclude that optical diffractometry of immunostained cells and Fourier analysis of cell shape are complementary to photomicroscopy for the study of the CMTC in cell populations cultured on an artificial substrate.

Alkaloids↗

PubMed1986

Fourier analysis of the cell shape of paired human urothelial cell lines of the same origin but of different grades of transformation.

The rationale of the present investigation is the observations made by many authors of changes in the molecular structure of the cell surface during the multistep process of malignant transformation. These changes may influence cell-matrix and cell-cell interactions and thereby cause changes in cell adhesiveness and cell shape. The aim of the present work was to investigate whether the development of various grades of transformation in vivo and in vitro of human urothelial cells is accompanied by significant changes in cell shape as measured by Fourier analysis. The following transformation grades (TGr) have been defined (Christensen et al. 1984; Kieler 1984): TGr I = nonmalignant, mortal cell lines that grow independently of fibroblasts and have a prolonged life span. TGr II = nonmalignant cell lines with an infinite life span. TGr III = malignant and immortal cell lines that grow invasively in co-cultures with embryonic chick heart fragments and possess tumorigenic properties after s.c. injection into nude mice. Comparisons of 4 pairs of cell lines were performed; each pair was of the same origin. Two pairs--each including a TGr I cell line (Hu 961b and Hu 1703S) compared to a TGr III cell line (Hu 961a or Hu 1703He)--were derived from two transitional cell carcinomas (TCC) containing a heterogeneous cell population. Two additional cell lines classified as TGr II (HCV-29 and Hu 609) were compared to two TGr III sublines (HCV-29T and Hu 609T, respectively) which arose by "spontaneous" transformation during propagation in vitro of the respective maternal TGr II-cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoma, Transitional Cell↗

PubMed1986

Objective evaluation of fibrosis in human testicular biopsies by analysis based on optical diffractometry.

Bilateral testicular biopsies were obtained from 27 patients and submitted to two different treatments for each of them: firstly, a classical fixation and colouring, permitting the histopathological diagnosis and subjective appreciation of the degree of fibrosis; secondly, a new objective technique based on optical diffractometry: this method is based on the analysis of light intensity distribution in the diffraction pattern of an original image. Using two different methods of discriminant analysis, we observed some errors due to the subjective examination; in particular, several fibrosed samples had been judged as 'normal' by the histopathologist. Moreover, we showed the existence of some heterogeneity between different slides from the same original biopsy.

Biopsy↗

PubMed1984

Fourier analysis of the shape of normal and transformed epithelial cells derived from human transitional epithelium.

The aim of this paper is to show the possibility of objective mathematical description of changes occurring in the shape of cells in the process of transformation. The evaluation of the changes in cell shape of the chosen cell lines differing in transformation grade was performed by the use of Fourier analysis of the shape. Any two-dimensional contour can be described with specific accuracy in a mathematical manner using the closed form Fourier series of cosines. The components forming the analysed shape, called harmonics, are independent and uncorrelated measures of their contribution to the total shape. The shape of each cell can be represented by the spectrum of harmonic amplitudes. To quote the paper by Healy-Williams and Williams (1981): "The observed shape is partitioned into series, where gross shape, as elongation or triangularity, is measured by the harmonic amplitudes of the lower harmonic order and increasingly fine scaled surface sculpture is measured at higher orders". The statistically evaluated results allow the objective comparison of the cell shapes of several compared cell lines differing in transformation grades. Malignant transformation is supposed to be a multistep process. The different grades of transformation could be defined by several parameters as changes in the morphology of the cells, their ability to compete with fibroblasts, their life span, their angiogenic potency, their invasiveness in vitro and their tumorigenicity in nude mice. In this paper several human urothelial cell lines of normal and tumor origin differing in their transformation grade (TGr I-III) were compared by the use of Fourier analysis of their shape. TGr I cultures have finite life span but do not need intermittent collagenase treatment to prevent fibroblast overgrowth. TGr II cultures acquire infinite growth potential, here defined as capacity to survive at least 70 passages. They are neither tumorigenic nor invasive. TGr III cultures show infinite growth transformation, increased angiogenicity and ability to invade normal host tissue in vitro. They produce progressively growing tumors in nude mice. The following human uroepithelial cell lines differing in the degree of transformation were studied and compared by statistical evaluation of the harmonic amplitudes describing mathematically the cell shape: Two cell lines derived from human transitional cell carcinoma (TCC): 1. Hu 1703S classified as TGr I, 2. Hu 1703He classified as TGr III. It was found that these two cell lines differ in all harmonics. Two cell lines derived from morphological normal human bladder epithelium: 3. HCV-29 classified as TGr II.(ABSTRACT TRUNCATED AT 400 WORDS)

Carcinoma, Transitional Cell↗

PubMed1984

Crystallinity of mineral deposited in arterial walls in the course of arteriosclerosis in diabetics and in patients with normal carbohydrate metabolism.

A comparison was made of the amount and crystallinity of mineral deposited in the course of the arteriosclerosis in arterial walls in diabetic patients and in individuals with normal carbohydrate metabolism. The macroscopically unchanged tunica intima, fibro-lipidic plaques and bone-like lamellae were taken from aorta thoracalis , aorta abdominalis, arteriae femorales and arteriae coronariae in the course of the autopsy of 17 insulin dependent diabetic individuals and of 9 persons with arteriosclerosis, but with normal carbohydrate metabolism, called control group. The total inorganic constituents in the three kinds of samples were determined by ashing of dried samples at 600 degrees C for 6 h. Crystallinity of mineral was defined as the ratio of the spins connected with the radiation-induced stable paramagnetic centers present in the crystalline lattice of hydroxyapatite crystal, to the total ash content of the sample. An increase in the amount and in the crystallinity of deposited mineral was observed when consecutive stadia of development of arteriosclerotic lesions were compared. This phenomenon resembles the maturation of mineral observed in the course of bone development.

Aged↗

PubMed1983

Application of optical diffractometry in studies of cell fine structure. Comparison of arterial smooth muscle cells in contractile and synthetic state.

Arterial smooth muscle cells in contractile and synthetic state were analyzed by optical diffractometry. Cell sections (80-90 nm) were photographed in an electron microscope and diffraction patterns of the plates (negatives) were produced using a helium-neon laser. Radial and angular distributions of light intensity in the diffractograms were measured and digitized using an electronic detector plate consisting of ring- and wedge-shaped photosensitive elements; radial distributions provide information about size of structures and distances between them and angular distributions about spatial orientation of structures in the images. Micrographs of nuclei and cytoplasm were analyzed separately (40-50 plates in each group). Computerized statistical analysis of radial distributions of light intensity showed that the nuclear chromatin pattern differed between cells in contractile and synthetic state. The probability that the observed difference could have arisen purely by chance was less than 10(-5). Computer-aided classification to the a priori known cell group was correct in 96.5% of the cases. Analysis of radial distributions of light intensity similarly showed marked differences in cytoplasmic structure between cells in contractile state (dominated by bundles of myofilaments) and synthetic state (dominated by cisternae of rough endoplasmic reticulum). The probability that the observed difference could have arisen purely by chance was less than 10(-5). Computer-aided classification to the a priori known cell group was correct in 92.0% of the cases. In contrast, analysis of angular distributions of light intensity did not indicate any statistically significant differences between contractile and synthetic state cells. A likely reason is that both myofilaments and cisternae of rough endoplasmic reticulum were arranged in parallel. The results demonstrate that optical diffractometry is a useful method for image analysis in studies of cell fine structure. It provides information about size and orientation of structures with poorly defined shape and is particularly well suited for studies on cell differentiation and effects of pharmacological and other experimental treatments on cell fine structure. It represents an alternative and a complement to stereology for quantitative and objective evaluation of morphological data.

Animals↗

PubMed1982

Application of the optical Fourier transform for analysis of the spatial distribution of collagen fibers in normal and osteopetrotic bone tissue.

Optical Fourier analysis was applied for evaluation of the differences between normal and pathologically changed bone tissue. Collagen fibers were used as markers of bone structure. To prove the usefulness of this technique for objective mathematical analysis of these differences the spatial spatial distribution of collagen fiber bundles was evaluated in normal and osteopetrotic bone. The variation in the spatial distribution of collagen fiber bundles in cross sections of femur diaphyses was evaluated quantitatively by optical diffraction three groups of Fatty Orl-op strain rats, i.e. phenotypically normal animals, osteopetrotic (op/op) mutants and op/op-mutants cured by transplantation of normal syngenic bone marrow. The histological sections of decalcified bone were stained with Sirus-Red and then photographed under polarizing microscope. The Sirus-Red staining was used to enhance the natural birefringency of collagen fibers. Diffractograms obtained from microphotographs of selected bone section areas, i.e. outer and inner circumferential lamellae and haversian bone of normal and cured op/op animals as well as whole cortical bone and woven bone filling the medullary cavities in op/op mutants were analysed separately. Diffractograms contain summarized information on the size and relative position of these structures in histological sections. The radial and angular distribution of light energy were evaluated for each diffractogram with an electronic detector. The obtained distributions were described by several sets of parameters concerning the position, level and shape of local maxima and minima. Out of these parameters five with the highest discriminant power were chosen for further mathematical analysis. This analysis was based on the calculation of the position of centroids in the multidimensional space described by the linear functions of the chosen parameters for each of the evaluated bone section areas. The centroids (mean values of discriminant scores of each group) represent the centers of gravity of the analysed groups, while the separation of the centroids tested by the F-test illustrates the differences between the respective groups of selected bone section areas. A high level of separation of centroids was found when osteopetreotic bone was compared with normal one, what means that the spatial distribution, size and interstructural distances between the collagen fiber bundles in bone tissue in these two groups of animals differ markedly. A similar situation was observed when osteopetrotic bone was compared with bone tissue obtained from op/op mutants cured by transplantation of normal syngenic bone marrow. On the other hand, the level of separation of centroids was low when bone tissue of cured op/op mutants was compared with the control one, a finding which corresponds to the less pronounced histological differences between these two groups of animals. Computer-aided classification on single microphotographs of selected bone section areas to the known a priori type of bone tissue was performed...

Animals↗

PubMed1982

Polarizing microscopy of Picrosirius stained bone sections as a method for analysis of spatial distribution of collagen fibers by optical diffractometry.

Cross sections of femur diaphysis obtained from control and osteopetrotic rats were stained with hematoxylin-eosin (HE) and Picrosirius (SR). Analogous selected areas of bone sections photographed under a polarizing microscope were analysed by optical diffractometry. Since the collagen fibers are a good marker for the structure of bone tissue, their spatial distribution evaluated by optical diffractometry provides information on the tissue architecture. The Picrosirius staining technique enhances the natural birefringency of collagen fibers. Therefore, in the polarizing microscope, pictures of high contrast are obtained. This procedure, by increasing the amount of information in the image, increases the quantity of the data obtained by optical diffractometry in comparison with the HE staining method. The results obtained prove that SR staining combined with polarizing microscopy might be useful for optical diffractometry in analysis of the spatial distribution of collagen fibers in all connective tissues, where they could serve as markers of tissue architecture.

Animals↗