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Biomedical subjects

J Aigner

Publications and source records attributed to J Aigner.

At least 19 recordsLinked to original sources

Effect of different gamma-irradiation doses on cytotoxicity and material properties of porous polyether-urethane polymer.

Biomaterials respond to sterilization methods differently. Steam sterilization might decrease the performance of thermoplastic polyether-urethane (TPU); however, the effect of different gamma-radiation doses on this polymer is contradictory in present literature. The purpose of this study was to investigate the differences between irradiative doses in comparison with steam sterilization on a porous TPU scaffold produced by a new processing method. No significant differences in the surface chemical structure were found with attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) analysis when comparing with the sterilization methods. The molecular weight (M(w)) had a net increase from 11.5 +/- 0.039 to 13.2 +/- 0.072 kDa by gamma-sterilization from 10 to 60 kGy. The samples that were irradiated (>60 kGy) had also an increase in polydispersity index (PDI; 1.45 +/- 0.007) in comparison with the nonsterile ones (1.31 +/- 0.017), which indicate branching. Liquid chromatography/mass spectroscopy (LC/MS) analysis showed that there was a correlation between the concentration of the breakdown product, methyl dianiline, and cytotoxicity. The concentration of this compound was found to be four times higher in steam-sterilized sample (1.3 +/- 0.01 ppb) compared with that of the polymer sample gamma-sterilized at 10 kGy (0.3 +/- 0.01 ppb). The cytotoxicity of TPU was found to decrease with higher radiation doses, and was significantly higher for the steam-sterilized samples. It is recommended that TPU produced with the described foaming method should be sterilized by gamma-irradiation at 25 kGy or higher doses.

Biocompatible Materials↗

A novel processing method for injection-molded polyether-urethane scaffolds. Part 2: cellular interactions.

A large-scale scaffold processing method with injection molding has been successfully developed. Water was used as a foaming agent for the new technique. NaCl was used as a porogen to achieve an open-cell structure. Organic solvents, which are common foaming agents for polyurethane, where not used. Toxic remains in the polymer were therefore prevented. Biocompatibility tested gave a mean optical density of 81% from WST-1 proliferation assay. In comparison to the previously study processing method, hot pressing (Haugen H, Ried V, Brunner M, Will J, Wintermantel E. J Mater Sci: Mater Med2004;15:343-346), the current scaffolds had an increase of 20% of the mean optical density. Cell seeding showed that human fibroblasts adhered to the surface and proliferated. The spread of the adhered fibroblasts was uniform on the surface. A quantitative MTT analysis proved that there was a significant (p < 0.01) increase in the OD level after 7 and 14 days of incubation. This cell layer thickened with increased incubation time from 7 to 14 days (p < 0.05) and had typical fibroblast morphology.

Biocompatible Materials↗

[Techniques for functional tissue and organ replacement using postnatal stem cells].

Postnatal stem cells play a decisive role in cell-based therapies due to their high proliferation activity and functional plasticity. On the one hand, basic research in cell biological processes of adult stem cells is crucial in order to establish them as therapeutic tools. On the other hand, development and enhancements of appropriate techniques are required: we need to establish defined technologies for extraction and differentiation of stem cells and to develop adequate cell carrier devices, scaffolds, and bioreactors for in vitro purposes. Furthermore, it is an interdisciplinary challenge to consider logistical aspects concerning isolation, transport, and storage of stem cells in order to use them in a wide range of activities in regenerative medicine. In this review we present the current methods of work and research on adult stem cells. We explain their therapeutic use and define requirements for future technological developments for work with postnatal stem cells.

Bioreactors↗

[The STEMMAT-project as part of health initiative BayernAktiv: adult stem cells from umbilical cord and cord blood as alternative to embryonic stem cell research].

Adult stem cells from umbilical cord and cord blood are an interesting alternative to embryonic stem cells because such research is commonly recognized as ethical undisputed and many aspects are still insufficiently investigated. In the context of the STEMMAT research project (STEM = Stem Cell and MAT = Material) different aspects of stem cells from umbilical cord and cord blood are investigated, to improve basic science understanding and potentially leading someday to a clinical application.

Cord Blood Stem Cell Transplantation↗

[Tissue-engineered cartilage in a prefabricated microvascularized flap].

INTRODUCTION: In reconstructive surgery, the integration of tissue-engineered cartilage in a prefabricated free flap may make it possible to generate flaps combining a variety of tissue components to meet the special requirements of a particular defect. The aim of the present study was to establish the technique of prefabricating a microvascular free flap by implanting a vessel loop under a skin flap in a rabbit model. The second aim was to gather experience with prelaminating the flap with autologous tissue-engineered cartilage in terms of matrix development, inflammatory reaction and host-tissue interaction. METHODS: The microvascular flap was created by implanting a vessel loop under a random pattern abdominal skin flap. The tissue-engineered cartilage constructs were made by isolating chondrocytes from auricular biopsies. Following a period of amplification, the cells were seeded onto a non-woven scaffold made of a hyaluronic acid derivative and cultivated for 2-3 weeks. One cell-biomaterial construct was placed beneath the prefabricated flap, and the others were placed subcutaneously under the abdominal skin and intermuscularly at the lower extremity. In addition, a biomaterial sample without cells was placed subcutaneously as a control. All implanted specimens were left in position for 6 or 12 weeks. After explantation, the specimens were examined by histological and immunohistological methods. The prefabricated flap was analyzed by angiography. RESULTS: The prefabricated flaps showed a well-developed network of blood vessels formed by neovascularization between the implanted vessel loop and the original random-pattern blood supply. The tissue-engineered constructs remained stable in size and showed signs of tissue similar to hyaline cartilage, as evidenced by the expression of cartilage-specific collagen type II and proteoglycans. No hints of inflammatory reactions were observed. CONCLUSION: These results show the potential of prefabricated flaps as custom-made flaps for reconstructive surgery in difficult circumstances, more or less independent of anatomical prerequisites. Cartilage tissue engineering provides a 3-dimensional structure with minimal donor-site morbidity.

Animals↗

[Umbilical cord stromal cells (UCSC). Cells featuring osteogenic differentiation potential].

The identification of appropriate cell types is necessary to establish cell-based therapies in regenerative medicine. These cell types must (1) be available in an appropriate amount, (2) be easy to obtain, (3) be sufficiently expandable in vitro, and (4) fit to or at least be able to differentiate into the required cell type. Since the umbilical cord is available without any intervention and represents a notable amount of tissue, we consider it to be a promising source for isolating cells for cell-based therapies. This study demonstrates that umbilical cord stromal cells (UCSC), the connective tissue cells of the umbilical cord, can be isolated in sufficient quantities and be well expanded. UCSC feature phenotypic plasticity and thus are functionally similar to stem cells. UCSC can be differentiated into cells with osteoblastic properties (expression of alkaline phosphatase, formation of bone nodules). It is concluded that the umbilical cord should no longer be regarded as valueless tissue and be unthinkingly discarded. Instead, it should be considered a valuable resource for the isolation of potent cells for cell-based therapies, especially for treatment of bone defects.

Alkaline Phosphatase↗

Clinical aspects and strategy for biomaterial engineering of an auricle based on three-dimensional stereolithography.

At the present time, the partial and/or complete reconstruction of an auricle from autologous rib cartilage is one of most widely published techniques. In the field of tissue engineering, different techniques have been described to generate cartilage tissue using isolated chondrocytes. The basis of these tissue-engineering techniques is bioresorbable or non-bioresorbable biomaterials, which serve as a three-dimensional cell carrier. Tissue engineering of an auricle requires preformed bioresorbable biomaterials designed to fit the form of a patient's auricular defect. Three-dimensional imaging acquired from computed tomography scans or laser surface scanning has become an important tool in modern medicine. This study represents the preoperative procedures for the reconstruction of an auricle through tissue engineering in accordance with the clinical aspects. Hyaff 11, a hyaluronic acid derivative, was used as a three-dimensional cell carrier for isolated human nasoseptal chondrocytes. The chondrocytes were amplified in a conventional monolayer culture before the cells were seeded on a hyaluronic non-woven mesh and cultured in vitro for 4 weeks. The chondrogenic potential of human nasal chondrocytes in Hyaff 11 was investigated by confocal laser scanning microscopy, histology (toluidine blue) and immunohistochemistry (collagen type II). Computer-aided design (CAD) and manufacture of an auricle model with stereolithographical methods were used for the prefabrication of a bioresorbable three-dimensional cell carrier designed in the form of a patient's auricular defect. The cell carrier used was Hyaff 11, a fully benzyl-esterified hyaluronic acid derivative. Confocal laser scanning microscopy has shown good cell attachment, a homogenous distribution of amplified chondrocytes and a viability of more than 90%. After 4 weeks in vitro culture the human nasoseptal chondrocytes synthesized new cartilage with the expression of cartilage-specific collagen type II. In order to shape a patient's designed scaffold the auricle model was fitted exactly and symetrically to the contralateral side. Subsequently, the mirror image patient-specific model was used to prepare an identical scaffold model made of a fully benzyl-esterified hyaluronic acid derivative. The bioresorbable scaffold that was produced gave a satisfactory representation of auricle structure. Bioresorbable preformed biomaterials in the form of a patient's auricle defect represent an important prerequisite for the tissue engineering of autologous auricle grafts. Hyaff 11 seems to be a promising material for tissue engineering of cartilage transplants, and the application of this approach will improve conventional reconstructive surgery in the future.

Absorbable Implants↗

[Mesenchymal stem cells--a new pathway for tissue engineering in reconstructive surgery].

INTRODUCTION: Mesenchymal stem cells (MSC) have the capacity to differentiate into chondrocytes with the synthesis of cartilage. This report presents the use of human adult bone marrow derived mesenchymal stem cells for tissue engineering of autologous cartilage grafts. METHODS: Human bone marrow aspirates were obtained from the iliac crest and fractionated on a Percoll gradient. The isolated hMSC were plated at 20 x 10 (6) cells per 100 mm (2) culture dish. After 21 days in culture at 37 degrees C with 5 % CO 2, the adherent multiplied MSC were trypsinized, counted, and tested for viability by trypan blue assay. The hMSCs were loaded into a sterile 15 ml polypropylene tube (0.5 Mio cells/ml) and centrifuged on the bottom of the tube at 500 g for 5 minutes. The MSC were cultivated for 3 weeks in vitro in a specific chondrogenetic medium composed of Dulbecco's Modified Eagles Medium-High Glucose supplemented with 10 ng/ml transforming growth factor-beta 1, 1 % ITS-Premix medium, 80 micro M ascorbic acid, and 100 nM dexamethasone. RESULTS: Histological and immunohistochemical studies performed after 3 weeks in three dimensional culture demonstrated the expression of cartilage specific collagen type II and X as well as proteoglycans. CONCLUSION: Human adult mesenchymal stem cells derived from bone marrow aspirates have the ability to differentiate into chondrocytes under specific culture conditions by growth factors. The use of adult mesenchymal stem cells may be a promising tool for tissue engineering of autologous cartilage grafts in reconstructive surgery in the future.

Adult↗

Screening for Aspergillus spp. using polymerase chain reaction of whole blood samples from patients with haematological malignancies.

Sensitive screening for Aspergillus spp. using polymerase chain reaction (PCR) of whole blood samples in patients with haematological disorders has not been performed to date. In a 2-year study, 121 patients admitted to the University Hospital of Innsbruck for cancer chemotherapy without clinical signs of fungal infection were prospectively screened for Aspergillus spp. In 28 out of 121 (23%) patients, Aspergillus DNAaemia was detected. Of these patients, 16 (57%) were positive only once for Aspergillus DNA, but positivity was never associated with invasive aspergillosis. PCR positive episodes were short and resolved without antifungal treatment. Five patients (18%) had intermittent PCR positive results. Seven (25%) patients presented at least two consecutive positive PCR results; one of these patients developed invasive aspergillosis and another two were strongly suspected as having aspergillosis. Based on the criteria of the European Organization for Research and Treatment of Cancer case definitions, sensitivity and specificity of serial PCR monitoring were 75% and 96%. Positive PCR results became negative shortly after commencement of antifungal treatment, but the changes did not correlate with clinical responsiveness to treatment in three patients. Our results indicate the potential usefulness of PCR for screening for Aspergillus spp. in patients at risk, but without antifungal treatment.

Adult↗

[Microtia: technique for external ear reconstruction with autologous rib cartilage].

Various methods for treatment of classic microtia are known. Beside a prosthesis, the most common way of auricle reconstruction is the use of autogenous rib cartilage; a process that requires two to three operations. In the first operation, rib cartilage is harvested from the 6th to the 9th rib. The base of the framework is the 6th and 7th rib cartilage which is taken under preservation of the synchondrosis. To mimic a 3-dimensional structure, the triangular fossa and scapha are carved into the groundplate and the 8th rib is fixed as a helical rim. After optimising the framework, it is placed in a subcutaneous pocket on the mastoid plane. In a second operation, approximately three months later, the auriculocephalic angle is reconstructed with a cartilage wedge, which is covered by a temporalis fascia flap and split skin-graft from the hairbearing skull. Commonly, a third operation is needed for minor refinements. Currently, autogenous rib cartilage is the ideal material available for ear reconstruction resulting in an excellent cosmetical outcome, although harvesting of the cartilage causes a specific donor-site morbidity. Operations improving the hearing ability by building up the external hearing channel and middle ear are mainly done in cases of bilateral microtia. Ear reconstruction with autogenous rib cartilage produces a replicable aesthetic result. The patients should be at least eight years old.

Adolescent↗

Relapsing polychondritis: a course over 20 years with cerebral involvement.

Relapsing polychondritis is a chronic inflammatory disease associated with an autoimmune disorder in cartilaginous tissue, eyes, labyrinth, blood vessels, and central nervous system. We describe a 75-year-old woman who presented with a 20-year history of dyspnea, inspiratory stridor, and polyarthritis. She developed dysmorphism of both ears and a saddle nose approximately 10 years earlier. Subsequently, she suffered from hearing loss and a tremor. A T2-weighted magnetic resonance imaging scan of the brain revealed multiple, spotted signal intensities. Immunohistochemical analysis of a serum sample showed antibodies to cartilaginous tissue, which were further identified on immunoblotting as antibodies to type II collagen. The extremely prolonged course of disease (>20 years) until a correct diagnosis was made is remarkable. Also, cerebral involvement, which was most likely caused by cerebral angiitis, and which, to our knowledge, has never previously been reported in this form, was detected. Arch Otolaryngol Head Neck Surg. 2000;126:1495-1498

Aged↗

[Highly malignant T-cell non-Hodgkin lymphoma (nasal type) of the hard palate].

The monomorphic clinical aspect of destructive mid-face lesions is characterised by inflammation, induration and granulomatous transformation. This feature can be caused by various infections, toxical noxa, Wegener's Granulomatosis and different neoplasms. The case of a 19 year old patient with EBV associated nasal type T-cell lymphoma located at the hard palate is presented. The diagnostic approach and difficulties in diagnosing this entity assessing by using multiple biopsies, serological and molecularbiological detection of EBV association and immunohistochemistry for atypic T-cells are elucidated. In the presented case the treatment with chemotherapy and irradiation following a well-defined therapy concept leaded to a three year recurrence-free survival so far. The comparison of the key-histological findings and the major differential diagnoses is mandatory to establish the final diagnosis of lymphoma. This is the basement for treating this disease with combined chemotherapy and irradiation for optimizing survival.

Adult↗

Tissue-engineered cartilage using serially passaged articular chondrocytes. Chondrocytes in alginate, combined in vivo with a synthetic (E210) or biologic biodegradable carrier (DBM).

In vitro multiplication of isolated autologous chondrocytes is required to obtain an adequate number of cells to generate neo-cartilage, but is known to induce cell-dedifferentiation. The aim of this study was to investigate whether multiplied chondrocytes can be used to generate neo-cartilage in vivo. Adult bovine articular chondrocytes, of various differentiation stages, were suspended in alginate at densities of 10 or 50 million/ml, either directly after isolation (P0) or after multiplication in monolayer for one (P1) or three passages (P3). Alginate with cells was seeded in demineralized bovine bone matrix (DBM) or a fleece of polylactic/polyglycolic acid (E210) and implanted in nude mice for 8 weeks. The newly formed tissue was evaluated by Alcian Blue and immunohistochemical staining for collagen type-II and type-I. Structural homogeneity of the tissue, composed of freshly isolated as well as serially passaged cells, was found to be enhanced by high-density seeding (50 million/ml) and the use of E210 as a carrier. The percentage of collagen type-II positive staining P3-cells was generally higher when E210 was used as a carrier. Furthermore, seeding P3-chondrocytes at the highest density (50 million/ml) enhanced collagen type-II expression. This study shows promising possibilities to generate structurally regular neo-cartilage using multiplied chondrocytes in alginate in combination with a fleece of polylactic/polyglycolic acid.

Alcian Blue↗

[Subcutaneous preservation of an amputated auricle. Morphological changes].

BACKGROUND: Microsurgical replantation of an avulsed auricle remains a challenge in reconstructive surgery. Secondary reconstruction of a traumatic lost auricle is usually performed using a costal cartilage framework according to well documented techniques or with a prosthesis. In order to minimize donor-site morbidity, various efforts can be undertaken to preserve the amputated auricle by implanting the de-epithelialized cartilage framework in a subcutaneous pocket on the surface of the mastoid. Where preservation is successful, this original cartilage could be used for reconstructive treatment. PATIENT AND RESULTS: This study describes the histologic and immunohistologic changes in a complete traumatic avulsion of the auricle with subsequent cartilage conservation for eight months within a skin pocket. Trauma, preparation and preservation were accompanied by morphologic changes that included generation of local ossification centers and infiltration of fibrous tissue. We compared the macroscopic and microscopic morphology of the amputated part to native elastic cartilage following maximal denutrition and temporary heterotopic implantation in conjunction with atypical tension and pressure properties of the retroauricular pocket. CONCLUSION: In this case, the limited success of cartilage conservation in the subcutaneous pocket required conventional auricle reconstruction with autologous costal cartilage.

Adult↗

Cartilage reconstruction in head and neck surgery: comparison of resorbable polymer scaffolds for tissue engineering of human septal cartilage.

New cell culture techniques raise the possibility of creating cartilage in vitro with the help of tissue engineering. In this study, we compared two resorbable nonwoven cell scaffolds, a polyglycolic acid/poly-L-lactic acid (PGA/PLLA) (90/10) copolymer (Ethisorb) and pure PLLA (V 7-2), with different degradation characteristics in their aptitude for cartilage reconstruction. Chondrocytes were isolated enzymatically from human septal cartilage. The single cells were resuspended in agarose and transferred into the polymer scaffolds to create mechanical stability and retain the chondrocyte-specific phenotype. The cell-polymer constructs were then kept in perfusion culture for 1 week prior to subcutaneous transplantation into thymusaplastic nude mice. After 6, 12, and 24 weeks, the specimens were explanted and analyzed histochemically on the presence of collagen (azan staining), proteoglycans (Alcian blue staining), and calcification areas (von Kossa staining). Furthermore, different collagen types (collagen type I, which is found in most tissues, but not in hyaline cartilage matrix; and collagen type II, which is cartilage specific) were differentiated immunohistochemically by the indirect immunoperoxidase technique. Vascular ingrowth was investigated by a factor VIII antibody, which is a endothelial marker. Quantification of several matrix components was performed using the software Photoshop. Significant differences were found between both nonwoven structures concerning matrix synthesis and matrix quality as well as vascular ingrowth. Ethisorb, with a degradation time of approximately 3 weeks in vitro, showed no significant differences from normal human septal cartilage in the amount of collagen types I and II 24 weeks after transplantation. Thin fibrous tissue layers containing blood vessels encapsulated the transplants. V 7-2 constructs, which did not show strong signs of degradation even 24 weeks after transplantation, contained remarkably smaller amounts of cartilage-specific matrix components. At the same time, there was vascular ingrowth even in central parts of the transplants. In conclusion, polymer scaffolds with a short degradation time are suitable materials for the development of cartilage matrix products, while longer stability seems to inhibit matrix synthesis. Thus, in vitro engineering of human cartilage can result in a cartilage-like tissue when appropriate nonwovens are used. Therefore, this method could be the ideal cartilage replacement method without the risk of infection and with the possibility of reconstructing large defects with different configurations.

Adult↗

Cartilage tissue engineering with novel nonwoven structured biomaterial based on hyaluronic acid benzyl ester.

The aim of this study was to investigate the possibility of using the benzyl ester of hyaluronic acid (HYAFF 11), a recently developed semisynthetic resorbable material, as a scaffold for the culture of human nasoseptal chondrocytes in tissue-engineering procedures of cartilage reconstruction. Different techniques such as immunohistochemistry, scanning electron microscopy, and confocal laser scanning microscopy were used to study the behavior, morphology, and phenotype expression of the chondrocytes, which were initially expanded and then seeded on the material. The nonwoven cell carrier allowed good viability and adhesivity of the cells without any surface treatment with additional substances. Furthermore, the cultured cells expressed cartilage-specific collagen type II, indicating that they were able to redifferentiate within the scaffold of HYAFF 11 and were able to retain a chondrocyte phenotype even after a long period of in vitro conditions. Nevertheless, the expression of collagen type I, which was produced by dedifferentiated or incompletely redifferentiated chondrocytes, was noticeable. Additional data were obtained by subcutaneous implantation of samples seeded with human cells in the in vivo model of the athymic nude mouse. The results after 1 month revealed the development of tissue similar to hyaline cartilage. This study is promising for the use of this scaffold for tissue engineering of cartilage replacements.

Animals↗

Transitional stages in the development of the rabbit renal collecting duct.

The collecting duct (CD) epithelium of the mammalian kidney is an extraordinary structure with respect to its functional changes during development and its heterogeneous composition when matured. All of the different nephron epithelia of the mammalian kidney consist of one single cell type. In contrast, the differentiated CD is composed of at least three distinct cell types [principal, alpha intercalated-, and beta intercalated cells] that are responsible for the multiple physiological functions of this kidney compartment. During development the function of the CD changes: initially, the CD ampulla serves as an embryonic inducer, while the matured epithelium plays a key role in maintaining the homeostasis of body fluids. At present the process of CD maturation is not well understood. Neither the time course of development nor the morphogenic factors leading to the heterogeneously composed epithelium are known. In the present study the differentiation of the CD epithelium was investigated using newly developed monoclonal antibodies and well-characterized antisera. The morphological changes induced during differentiation were monitored by immunohistochemistry and scanning electron microscopy. The experiments were performed on neonatal and adult rabbit kidneys. Results obtained by light microscopical techniques and scanning electron microscopy revealed that the ampullary tip can be distinguished from the ampullary neck, as well as from the maturing CD. A number of proteins that were not detectable in the ampulla were detected in the neonatal CD and were found at even higher concentrations in the adult CD (PCD8, chloride/bicarbonate exchanger). Other proteins (PCD9) were downregulated during differentiation. For the first time the transient character of the differentiation stage of the neonatal CD could be demonstrated unequivocally. Furthermore, considerable heterogeneity in protein expression patterns (PCD6 and PCD9) was demonstrated within the beta IC cell population of the mature CD.

Aging↗