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Biomedical subjects

G Kratz

Publications and source records attributed to G Kratz.

31 records · Page 2Linked to original sources
PubMed1996

Characterization of a new in vitro model for studies of reepithelialization in human partial thickness wounds.

Reepithelialization of artificial partial thickness wounds made in biopsies of human skin was determined after 3, 5, or 7 d of incubation, submerged or elevated to the air-liquid interface. The biopsies were reepithelialized within 5-7 d, with a more complete epidermal healing in wounds exposed to air. Both types of wounds showed similar time-course in deposition of basement membrane components, as detected by immunofluorescence labeling. Laminin and collagen type VII were deposited underneath the migrating tips, whereas collagen type IV was detected after reepithelialization. Markers of terminal differentiation showed a pattern close to normal in the air-liquid incubated wounds after reepithelialization. Involucrin was detected in the suprabasal regions of the migrating epidermis and thereafter in the upper half of neo-epidermis in the air-liquid incubated wound. Filaggrin could not be detected in the submerged wounds at any time during healing, whereas wounds exposed to air showed a well-differentiated epidermis by Day 7. Tritiated thymidine-incorporation indicated proliferation of epidermal and dermal cells during reepithelialization and a maintained viability, as shown by cultivation of endothelial- and fibroblast-like cells obtained from the dermis 7 d after wounding. Reepithelialization in this human in vitro model is supported by a matrix close to normal with the possibility of extracellular influences and cell-cell interactions and, in addition, the technique is simple and reproducible. Therefore, we suggest this model for studies of regeneration in culture and as a complement to in vivo studies on epidermal healing.

Basement Membrane↗
PubMed1995

Collagenase expression is rapidly induced in wound-edge keratinocytes after acute injury in human skin, persists during healing, and stops at re-epithelialization.

Collagenolytic activity has been reported previously in association with wounds. We used in situ hybridization and immunohistochemistry to localize cellular sites of interstitial collagenase production in acute wounds in human skin at days 1, 2, 4, 6, 9, and 14 after wounding. In vivo, collagenase expression peaked in migrating basal keratinocytes at the wound edge at day 1, then gradually decreased and was undetectable at day 9 when healing was complete. To minimize the effects of crust formation and inflammation, we examined the healing of wounds made with a 3-mm punch in organ-cultured skin. In these in vitro wounds, re-epithelialization occurred by 5-7 d in 10% serum, although remodeling of the connective tissue was minimal. Collagenase expression showed a similar pattern as in the in vivo wounds; it was detected in migrating keratinocytes already 4-6 h after wounding, peaked at 12-24 h, gradually decreased during the next few days, and subsided upon re-epithelialization. In dermal fibroblasts, on the other hand, expression of collagenase started considerably later, after 5-7 d in culture, and persisted after complete re-epithelialization, indicating that collagenase is differentially regulated in different cell types. Our findings also show that collagenase induction in keratinocytes does not require inflammation and occurs as a rapid response to wounding, suggesting that interstitial collagenase is not only necessary for remodeling of the extracellular matrix, but may also have a role in initiating migration of keratinocytes in wound healing.

Animals↗
PubMed1995

Keratinocyte conditioned medium stimulates type IV collagenase synthesis in cultured human keratinocytes and fibroblasts.

We have previously shown that conditioned medium from cultured human keratinocytes stimulates proliferation of a variety of cell types involved in wound healing, as well as re-epithelialization of wounds in human skin in vitro. We now present evidence for an autocrine/paracrine control of the synthesis of type IV collagenases in human keratinocytes and fibroblasts. During wound healing, keratinocytes migrate over the wound bed, an activity coupled with lysis of basement membranes, and hence requiring the presence of collagenases. Collagenases are also needed for the production and remodelling of the granulation tissue. In order to study the autocrine/paracrine control of collagenase production in keratinocytes and fibroblasts, we stimulated these cells in culture with conditioned medium from cultured keratinocytes. Protease synthesis was determined by affinity labelling with 3H-diisopropylfluorophosphoridate (DFP) and by zymography. Keratinocyte-conditioned medium was found to increase the expression of 72 and 92 kDa type IV collagenase in human keratinocytes, and the 72 kDa collagenase in human fibroblasts, indicating that an autocrine/paracrine control mechanism is involved in collagenase production in these cell types during wound healing. This increased expression of collagenases could be partly responsible for the stimulated healing seen in wounds treated with sheets of cultured keratinocytes.

Cells, Cultured↗
PubMed1994

Insulin like growth factor-1 and -2 and their role in the re-epithelialisation of wounds; interactions with insulin like growth factor binding protein type 1.

Insulin like growth factor (IGF) 1 and 2 which are present and actively synthesised in the wound fluid stimulate several cell types involved in the process of wound healing. To investigate the role of IGF-1 and 2 and in addition, the association between IGF and their carrier proteins, IGF binding proteins (IGFBP), we have used a newly established model for human wound healing in fresh biopsy material. Histological examination shows that IGF-1 stimulates efficient reepithelialisation of the wounds both alone and in the presence of recombinant IGFBP-1. In contrast, IGF-2 stimulates healing only when used in combination with IGFBP-1. These findings suggest that the two IGFs and their carrier proteins may function during different phases of wound healing and that both IGF-1 and 2 act as potent inducers of wound healing; this may have direct clinical implications.

Carrier Proteins↗
PubMed1992

Effect of recombinant IGF binding protein-1 on primary cultures of human keratinocytes and fibroblasts: selective enhancement of IGF-1 but not IGF-2-induced cell proliferation.

The present report describes the mitogenic effect of recombinant IGF-2 on cultured human keratinocytes and fibroblasts compared to that of IGF-1. Furthermore, the modulating effect of a recently expressed recombinant form of placental-derived IGF-binding protein 1 (IGFBP-1) on IGF-induced proliferation was examined. A dose-dependent increase, up to 100%, in cell proliferation was seen in cultured human keratinocytes with IGF-2 and -1 and the proliferative response was comparable to the effect of epidermal growth factor (EGF). In human fibroblasts, IGF-1 stimulated DNA synthesis up to 300% for IGF-1 and up to 200% for IGF-2. The mitogenic effect of IGF-1 was enhanced by IGFBP-1 in both cell types. In contrast, the IGF-2-induced mitogenic effect was unperturbed. These findings indicate that the interaction between IGFs and their binding proteins may induce different responses depending upon the ligand and the target cell.

Blotting, Western↗
PubMed1992

[Arthritic changes in small and large joints in digital and conventional x-ray images].

In order to evaluate the suitability of digital radiography for the diagnosis of skeletal abnormalities, inflammatory joint changes were used to test this technique. Early erosive changes, fine irregularities and marginal lamellar abnormalities demand a high degree of resolution from the imaging method. 6 observers studied images of the hand of 67 and of the foot of 19 patients. Joint changes were staged according to the scheme proposed by Larsen; in this study the early stages predominated. With similar techniques there were no significant differences between the digital and conventional techniques. Images of 39 large joints (knee, shoulder, elbow, ankle) were compared only subjectively since even minor differences in projection were able to obscure some erosions. However, in this case also both techniques appear to be of similar value.

Adult↗
PubMed1991

Conditioned medium from cultured human keratinocytes has growth stimulatory properties on different human cell types.

Evidence for growth-stimulatory properties of keratinocyte-conditioned medium (KCM) on human fibroblasts, endothelial cells, keratinocytes, smooth muscle cells, and a mouse fibroblast cell line (3T3 cells) is presented. On human fibroblasts KCM caused an increase of over 400% in DNA synthesis as revealed by 3H-thymidine incorporation and autoradiography. The proliferative effect was comparable to that of platelet-derived growth factor (PDGF), but was not inhibited by PDGF antibodies and exceeded that of transforming growth factor-alpha (TGF-alpha), epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I), and basic fibroblast growth factor (bFGF). Furthermore, KCM was found to stimulate smooth muscle cells, keratinocytes, and endothelial cells more potently than PDGF, EGF/TGF-alpha, and bFGF, respectively. KCM was also potent in stimulating thymidine incorporation in 3T3 cells, whereas EGF showed a twenty-fold weaker stimulatory effect. Because keratinocytes have been shown to secrete TGF-alpha, which binds to the EGF receptor, binding of factors in KCM to the EGF receptor was assayed. The displacement of radiolabeled EGF by KCM corresponded to a low concentration of EGF (0.5 ng/ml), implying that the growth-stimulatory effect of KCM was not mediated via activation of EGF receptors. Taken together, these results suggest the presence of hitherto unidentified growth-stimulatory factor(s), expressed and secreted by cultured human keratinocytes.

3T3 Cells↗
PubMed1990

Individualized adaptation of clothes for impaired persons. A comparison of two groups with and without experience of adapted clothes.

Impaired persons often have difficulties in obtaining suitable clothing. Questionnaires on this subject were sent to a consecutive series of impaired persons. Group 1 (n = 50) had received adapted clothes and group 2 (n = 81) had not. Questionnaire A (both groups) contained general questions on clothes and questionnaire B (Group 1) contained questions on adapted clothes. The result of the study shows a significant difference (p less than 0.01) between the need for adapted clothing in group 1 and group 2. No significant difference was found between the groups regarding the reason for obtaining and using adapted clothes. It was difficult for 94% in group 1 and 85% in group 2 to obtain clothes which were suitable for the individual's impairment and which made their daily activities easier. In group 1 the adapted garments were of most value when dressing and undressing (86%). They also reduced the time needed, by 31% for the individual, by 43% for a helper; and by 62% when going to the toilet. The adapted garments were considered smarter and more comfortable by 64%. Eighteen per cent considered that wearing them had increased their self-confidence.

Activities of Daily Living↗
PubMedDate not supplied

Dose-dependent hyperbaric oxygen stimulation of human fibroblast proliferation.

Diabetic wounds are characterized by a prolonged wound healing process with insufficient formation of granulation tissue. Systemic hyperbaric oxygen therapy has been observed to improve the healing of these wounds. However, the mechanism(s) responsible for these findings are not yet fully elucidated. In the present study we have studied the in vitro effects of hyperbaric oxygen on proliferation of human fibroblasts from normal skin and from chronic foot ulcers in non-insulin-dependent diabetics. A 1-hour exposure to hyperbaric oxygen at oxygen pressures between 106 and 300 kPa (795 to 2250 mm Hg) increased the proliferation in both diabetic and normal fibroblasts. The stimulatory effect was dose-dependent, with a peak increase in cell proliferation at 250 kPa and 200 kPa for normal and diabetic cells, respectively. The effects were not due to hydrostatic pressure per se. These results suggest that hyperbaric oxygen could stimulate fibroblast activity in the diabetic wound, a finding that could explain the enhanced formation of granulation tissue seen clinically in wounds treated with hyperbaric oxygen. We also speculate that mechanisms other than just increased oxygen availability may be responsible for our findings.

Journal Article↗
PubMedDate not supplied

Tissue expression of transforming growth factor-beta1 and transforming growth factor-alpha during wound healing in human skin explants.

In the dynamic and complex process of wound healing, locally produced growth factors are important mediators, although their actual roles have not been fully established. In the present study, the presence of transforming growth factor-beta1 and -alpha during the re-epithelialization of full-thickness wounds was investigated in an in vitro model of wound healing in human skin. The amounts of transforming growth factor-beta1 and -alpha secreted from the wound area were measured with enzyme immunoassays, and immunohistochemistry was used to study the localization of these two growth factors in the healing wound. The wounds were followed until they were completely re-epithelialized. The results showed a continuous increase in secreted transforming growth factor-beta1 throughout the re-epithelialization phase of healing followed by a decrease after its completion. The keratinocytes migrating out from the wound edges showed intense staining for transforming growth factor-beta1 which declined to the level of the surrounding epidermis after the wound was covered by a new epidermis. After the skin was wounded, a decrease both in secreted transforming growth factor-alpha and in immunostaining for this growth factor was apparent. Even though a minor increase in the immunoreactivity for transforming growth factor-alpha occurred after the completion of re-epithelialization, no increase in secreted transforming growth factor-alpha could be detected by enzyme immunoassay. These data suggest that keratinocytes modulate their expression of transforming growth factor-beta1 and -alpha during the wound healing process in human skin and that these changes may be controlled in part by autocrine pathways.

Journal Article↗
PubMedDate not supplied

Melanocytes in cultured epithelial grafts are depleted with serial subcultivation and cryopreservation: implications for clinical outcome.

Patchy hypopigmentation often occurs unpredictably in the skin regenerated from cultured epidermal autografts, especially when that skin is grown from frozen cells, serially passaged, or both. The impact of serial subcultivation and cryopreservation on melanocyte viability in the cultured epidermal autograft culture system was investigated. Serial subcultivation of human keratinocytes through as many as eight passages was performed, and melanocyte densities in confluent cultures at each passage were determined after specific labeling of melanocytes. The experimental cells were frozen before cultivation and between passages to determine the effect of standard cryopreservation on melanocyte survival. Freshly passaged cells that had not been frozen served as controls. Melanocytes were gradually depleted during fresh passage of epidermal cells but persisted through as many as seven passages. Freezing before or after the first passage or between subsequent passages resulted in a complete loss of melanocytes by the third or fourth passage. The findings suggest that cryopreservation should be avoided during cultured epidermal autograft production to optimize melanocyte survival and minimize pigmentation abnormalities that may occur after grafting.

Adult↗