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Elof Eriksson

Publications and source records attributed to Elof Eriksson.

17 recordsLinked to original sources
PubMed2007

Gene therapy in cutaneous wound healing.

Medical treatment of complicated acute and chronic wounds represents a persistent and increasing medical and economic problem in our health-care system. In this review, we discuss the potentials and limitations of current and future gene therapy for the treatment of complicated, acute and chronic wounds. Chronic non-healing wounds result in significant morbidity, prolonged hospitalization, lost time from work and enormous health-care expenses. There are constant efforts to improve the therapeutic modalities to local treatment of wounds. One of them is gene-therapy where the delivery of peptides directly into the wound provides a relatively new and exciting possibility. The two groups of peptides of particular interest are growth-factors and anti-microbial peptides. Gene delivery of these peptides provides not only the possibility of more targeted local delivery but also larger concentrations. Many new techniques for gene delivery to wounds have been developed in recent years. The combination of mechanical and viral or chemical vectors appear to have the greatest yield. This review provides an update on gene delivery to cutaneous wounds.

Animals↗
PubMed2006

A novel titanium wound chamber for the study of wound infections in pigs.

In the face of emerging multidrug-resistant microbes, reliable animal models are needed to study potential new therapies in infected wounds. To this end, we implanted screw-top titanium chambers subdermally in full-thickness wounds on both flanks (n = 6 per flank) of 2 Goettinger minipigs. After 1 wk, chambers were inoculated with Staphylococcus aureus, Pseudomonas aeruginosa, or vehicle only. Throughout the study, wound fluid was harvested for quantitative bacterial cultures to monitor infection. Animals were followed for 4 wk, after which tissue biopsies were taken for histologic analysis and quantitative bacterial counts. The implanted titanium chambers were well tolerated by the pigs throughout the study. After inoculation of the chambers, wound infection was established and maintained for 14 d. Despite infection, no systemic effects were noted. Cross-contamination was negligible, compared with the vehicle-only control. After tissue ingrowth, each chamber creates a closed system that allows harvest of exudate or application of substances without loss of material from the chamber. Because 12 chambers are implanted in each pig, researchers have the opportunity to compare multiple treatment options (for example, antibiotics, antimicrobial peptides, gene therapy) in the same animal, with no interindividual variation. We conclude that the use of titanium chambers in pigs provides a reliable and reproducible in vivo model to investigate wound healing, wound infection, and treatment options.

Animals↗
PubMed2004

Wet wound healing: from laboratory to patients to gene therapy.

Wet treatment of wounds has been used as an "irrigation" method since the seventh century. We have developed the concept of an in vivo tissue culture that facilitates wound healing and allows tissue engineering. A transparent, flexible, round chamber provides the wet environment. This system heals clean wounds as fast or faster than any other method, with less scarring. It allows delivery of analgesics, antibiotics, growth factors, growth media, and cells into the chamber, becoming a platform for tissue engineering. Gene therapy of the wound can be done in the chamber with growth factor and other genes. A tetracycline switch allows precise timing and amounts of expression and provides the opportunity for sequential expression of genes delivered at the same time.

Epidermal Growth Factor↗
PubMed2004

Gene therapy in the treatment of lower extremity wounds.

This article presents a brief overview of the etiology of chronic wounds of the lower extremities and their current medical and surgical treatment. Gene therapy as a potential tool for treating therapeutically challenging wounds is described in terms of the vectors employed in gene transfer, as well as the strategies used to promote wound healing. Results from animal model studies, as well as clinical trials, are presented.

Journal Article↗
PubMed2003

Gene therapy in wound healing.

Gene therapy is a new and emerging technology that has been catalyzed by the progress of the Human Genome Project. It employs the process of manipulating genes to achieve a clinically beneficial alteration in gene product. Wound healing lends itself to the application of gene therapy by virtue of the vast array of proteins involved in its complex cascade. This article provides an overview of the background to gene therapy and describes current techniques in use as applied to wound healing. The authors show the potential role that many candidate genes may offer in the future for optimizing wound healing through gene therapy.

Genetic Therapy↗
PubMed2003

Wound fluid bacterial levels exceed tissue bacterial counts in controlled porcine partial-thickness burn infections.

In the present study, an established controlled burn wound model was used to test the hypothesis that controlled surface contamination with is capable of generating a noninvasive method for the creation of a reproducible deep tissue burn wound infection. Using a liquid tight-wound chamber in Yorkshire pigs, partial-thickness burns were inoculated with saline-immersed for 24 hours. Noninoculated burns and unwounded skin immersed in normal saline served as controls. Bacterial cultures of wound fluid were performed daily, and tissue biopsies for bacteriological and histological evaluations were performed on days 1, 3, and 5. was only recovered from -inoculated wounds (tissue and fluid), whereas all controls contained endogenous only. The number of colony-forming units per gram of wound tissue did not correlate with the bacterial counts found in the overlying wound fluid for any wounds. Fluid counts were consistently higher than tissue counts by two logs. -inoculated wounds showed three times deeper tissue destruction than control wounds. Obtaining consistently deep tissue colonization without cross-contamination among wounds, this study introduces a noninvasive model for controlled burn wound infection suitable for future investigations regarding the efficacy of topical antibiotic wound treatment in experimental burns.

Animals↗
PubMed2002

Cultured autologous fibroblasts augment epidermal repair.

BACKGROUND: Autologous dermal fibroblasts may be useful in the treatment of skin wounds and for the enhancement of keratinocyte proliferation. This paper addressed the following questions: (1) can cultured fibroblasts (CF) be transplanted as suspensions to full-thickness skin wounds and do they influence wound healing; (2) will the transplanted CF be integrated into the new dermis; (3) can a transgene that encodes a secretable marker, human epidermal growth factor (hEGF), be expressed in the wound fluid by the transplanted CF; and (4) do CF cotransplanted with cultured keratinocytes (CK) influence the rate of wound healing? METHODS: Suspensions of CF were transplanted alone or together with CK to full-thickness wounds covered with liquid-containing chambers in an established porcine model. RESULTS: Transplantation of CF accelerated reepithelialization as determined from wound histologies and sequential measurements of protein efflux over the wound surface. CF transfected with a marker gene, beta-galactosidase, resulted in in vivo gene expression and demonstrated that transplanted CF integrated into the developing dermis. Transplantation of hEGF gene-transfected CF resulted in significant hEGF expression in wound fluid. The hEGF levels peaked at day 1 (2450 pg/ml) and then sharply decreased to low levels on day 6. CF cotransplanted with CK led to greater number of keratinocyte colonies in the wound and accelerated reepithelialization as compared with CK alone. CONCLUSIONS: Transplanted CF integrated into the dermis, accelerated reepithelialization, and improved the outcome of CK transplantation. CF may also be used for the expression of transgenes in wound and wound fluid.

Animals↗
PubMed2002

Autologous skin transplantation: comparison of minced skin to other techniques.

BACKGROUND: Skin grafting may be necessary to close nonhealing skin wounds. This report describes a fast and minimally invasive method to produce minced skin suitable for transplantation to skin wounds. The technique was evaluated in an established porcine skin wound healing model and was compared to split-thickness skin grafts and suspensions of cultured and noncultured keratinocytes. MATERIALS AND METHODS: The study included 90 wounds on 3 pigs. Fluid-treated full-thickness skin wounds were grafted with minced skin, split-thickness skin grafts, noncultured keratinocytes, or cultured keratinocytes. Controls received either fluid or dry treatment. The wound healing process was analyzed in histologies collected at Days 8 to 43 postwounding. Wound contraction was quantified by photoplanimetry. RESULTS: Wounds transplanted with minced skin and keratinocyte suspension contained several colonies of keratinocytes in the newly formed granulation tissue. During the healing phase, the colonies progressed upward and reepithelialization was accelerated. Minced skin and split-thickness skin grafts reduced contraction as compared to keratinocyte suspensions and saline controls. Granulation tissue formation was also reduced in split-thickness skin-grafted wounds. CONCLUSIONS: Minced skin grafting accelerates reepithelialization of fluid-treated skin wounds. The technique is faster and less expensive than split-thickness skin grafting and keratinocyte suspension transplantation. Minced skin grafting may have implications for the treatment of chronic wounds.

Animals↗
PubMed2002

Inhibition of herpes simplex virus type 2 (HSV-2) viral replication by the dominant negative mutant polypeptide of HSV-1 origin binding protein.

UL9-C535C, the trans-dominant negative mutant polypeptide of herpes simplex virus type 1 (HSV-1) UL9 origin binding protein, is a potent inhibitor of HSV-1 viral DNA replication. This study focused on testing whether HSV-1 UL9-C535C and a genetically engineered UL9-C535C-encoding HSV-1 recombinant virus CJ83193 could inhibit herpes simplex virus type 2 (HSV-2) infection. First, a stable cell line, R-C535C, expressing a high level of UL9-C535C in the presence of tetracycline and little or no UL9-C535C in the absence of tetracycline was established. The single step growth experiment showed that like HSV-1, the de novo synthesis of HSV-2 could be suppressed approximately 1000-fold by UL9-C535C expressed in R-C535C cells in the presence of tetracycline. Secondly, compared with cells singly infected with HSV-2, co-infection of Vero cells with HSV-2 and CJ83193 reduced the replication efficiency of HSV-2 in co-infected cells by 30-40 fold in a single-step growth assay, which coincided with marked reduction in viral late gene expression, but not the expression of viral immediate-early genes. Taken together, in view of our recent demonstration that CJ83193 can serve as an effective vaccine in preventing HSV-1 infection in mice, one can generate a CJ83193-like HSV-2 recombinant virus that could potentially function as a new therapeutic class of recombinant viral vaccine against HSV-2 infection.

Animals↗
PubMed2002

Comparative efficacy of the Schistosoma mansoni nucleic acid vaccine, Sm23, following microseeding or gene gun delivery.

Sm23 is an integral membrane protein expressed widely in the human parasitic worm Schistosoma mansoni. Sm23 has already been shown to elicit protective immune responses following immunization with peptides or DNA constructs. In this study, we evaluated the immunogenicity and the protective efficacy of the Sm23 DNA vaccine using two different intradermal DNA delivery methods: microseeding and gene gun. Using both techniques, all mice immunized with the Sm23-pcDNA construct generated Sm23-specific immunoglobulin (Ig)G antibody, while mice immunized with the control plasmid, pcDNA, did not. Antibody isotypes analysis revealed that microseeding elicited mainly IgG2a and IgG2b antibodies, with relatively low levels of IgG1 and IgG3. The relative IgG1/IgG2a ratio was 0.03, indicative of a Th1 type immune response. In contrast, gene gun immunization resulted in significantly higher levels of IgG1 and IgG3. The relative IgG1/IgG2a ratio in this case was 11, indicative of a Th2 type immune response. No significant difference in the levels of IgG2b was observed. Coimmunization with plasmid DNA encoding either interleukin (IL)-12 or IL-4 by microseeding did not affect the levels of IgG1, while the levels of IgG2a and IgG2b were reduced. On the other hand, the levels of IgG3 were significantly increased by IL-4, but unchanged by IL-12. Importantly, in all experiments, the Sm23-pcDNA vaccine provided statistically significant levels of protection against challenge infection. Microseeding immunizations resulted in higher levels of protection (31-34% protection) than gene gun immunization (18% protection). This suggests that the Th1 type immune response elicited by microseeding immunization was responsible for the higher protection levels. However, the protective effect of the vaccine was not affected by coadministering plasmids encoding either IL-12 or IL-4 using the microseeding technique.

Animals↗
PubMed2002

Wet wound healing.

Wound treatment in a flexible transparent chamber attached to the perimeter of the wound and containing a liquid has been extensively tested in preclinical experiments in pigs and found to offer several advantages. It protects the wound; the liquid medium or saline in the chamber provides in vivo tissue culture-like conditions; and antibiotics, analgesics, and various molecules can be delivered to the wound through the chamber. The wound chamber causes no injury to the wound itself or to the surrounding intact skin. Topical delivery of, for instance, antibiotics can provide very high concentrations at the wound site and with a favorable direction of the concentration gradient. A series of 28 wounds in 20 patients were treated with a wound chamber containing saline and antibiotics. Most patients had significant comorbidity and had not responded to conservative or surgical management with débridement and delayed primary closure or skin grafts. Six wounds had foreign bodies present; four of these were joint prostheses. Seven patients were on corticosteroids for rheumatoid arthritis, lupus, or chronic obstructive pulmonary disease, and four patients had diabetes. Most patients were treated with the wound chamber in preparation for a delayed skin graft or flap procedure, but one was treated with a wound chamber until the wound healed. Twenty-five of the wounds (89 percent) healed, and five wounds (18 percent) required additional conservative management after the initial chamber treatment and grafting procedure. Of the three wounds that did not heal, one healed after additional chamber treatment, one had a skin graft that did not take, and one required reamputation at a higher level. Antibiotic delivery was less than one intravenous dose daily, which avoided the potential for systemic absorption to toxic levels. Antibiotics such as vancomycin and gentamicin could be used in concentrations of up to 10,000 times the minimal inhibitory concentration. Forty-eight hours after application, 20 percent or more of the original antibiotic concentration was present in the wound chamber fluid. In conclusion, the wound chamber provides a safe, powerful tool in the treatment of difficult infected wounds.

Adolescent↗
PubMed2002

Gene therapy in soft tissue reconstruction.

Gene therapy is defined as the introduction of a therapeutic gene into a cell, whose expression can lead to a cure of a disease or offer a transient advantage for tissue growth and regeneration. The delivery of genes can be undertaken for a number of purposes, usually it is attempted to enhance or add a function to a cell or a tissue or to delete or reduce another function. In this brief overview we describe various vehicles and techniques that have been developed to deliver therapeutic genes into cells, such as viral vectors and physical/chemical gene delivery methods including naked DNA and particle-mediated gene transfer, the microseeding technique and the application of lipids. Furthermore we review the potential utility of gene therapy from the perspective of a reconstructive surgeon. Several tissues will be discussed, particularly muscle, tendon, nerve, bone, skin and wounds.

Cell Culture Techniques↗
PubMedDate not supplied

In vivo gene delivery of Ad-VEGF121 to full-thickness wounds in aged pigs results in high levels of VEGF expression but not in accelerated healing.

We have previously reported that endogenous vascular endothelial growth factor (VEGF) concentration in older pig wounds peaked later and at one-fourth the level of young pigs. These data suggested that VEGF might play a major role in the healing of full-thickness wounds in the aged pig. By in vivo gene transfer using the microseeding technique, we treated full-thickness wounds with different doses of VEGF-expressing adenoviral vector (Ad-VEGF) varying from 1 x 10(7) to 2.7 x 10(11) particles per wound (ppw). We found that the VEGF expression in wound fluid followed a dose-response pattern. However, when wounds were microseeded with the highest concentration of Ad-VEGF (2.7 x 10(11) ppw), diminished healing rates were found. We then determined the minimal functional concentrations of Ad-VEGF. We used five aged Yucatan minipigs, all retired breeders, to analyze the role of over-expression of 1 x 10(8) and 1 x 10(9) ppw of Ad-VEGF (n= 78) in terms of healing of full-thickness wounds, all 2.5 x 2.5 x 1 cm in size (n= 158). The Ad-VEGF solutions were delivered to the wound floor and borders by in vivo microseeding. Control wounds (n= 80) were microseeded with Ad-Lac-Z (n= 25), treated with saline (n= 49) or treated dry (n= 6). All wounds except for the dry-treated ones were covered with a wound chamber and a wet environment was created by injecting 2.5 ml saline into the chamber. Peak VEGF expression (2300-4000 pg/ml) was detected on days 2 or 3 post gene delivery. This level of VEGF expression was not seen in the saline (n= 49) or Ad-null (n= 25) control groups. The VEGF expression in wounds treated with 1 x 10(8) and 3 x 10(8) ppw (n= 39) exhibited a slower onset with a peak concentration of 400-920 pg/ml on days 5-7. Although high levels of VEGF expression were achieved in the local wound environment, we could not show a significant increase in neovascularization as compared to saline-treated wounds. No significant differences were observed in the rate of reepithelialization and wound contraction among groups of full-thickness wounds treated with Ad-VEGF, Ad-null mutant, or saline in the aged "wet wound healing" pig model. These results indicate that increased levels of VEGF in wounds produced by in vivo gene transfer have little effect on the healing of full-thickness wounds in the aged pig model. Moreover, significantly higher levels of VEGF expression by Ad-VEGF could lead to impaired wound healing.

Adenoviridae↗
PubMedDate not supplied

Adenoviral gene delivery to primary human cutaneous cells and burn wounds.

The adenoviral transfer of therapeutic genes into epidermal and dermal cells is an interesting approach to treat skin diseases and to promote wound healing. The aim of this study was to assess the in vitro and in vivo transfection efficacy in skin and burn wounds after adenoviral gene delivery. Primary keratinocytes (HKC), fibroblasts (HFB), and HaCaT cells were transfected using different concentrations of an adenoviral construct (eGFP). Transfection efficiency and cytotoxicity was determined up to 30 days. Expression was quantified by FACS analysis and fluorimeter. Cytotoxicity was measured using the trypan blue exclusion method. 45 male Sprague Dawley rats received 2x10(8) pfu of Ad5-CMV-LacZ or carrier control intradermally into either superficial partial thickness scald burn or unburned skin. Animals were euthanized after 48 h, 7 or 14 days posttreatment. Transgene expression was assessed using immunohistochemistry and bioluminescent assays. The highest transfection rate was observed 48 h posttransfection: 79% for HKC, 70% for HFB, and 48% for HaCaT. The eGFP expression was detectable in all groups over 30 days (P>0.05). Cytotoxic effects of the adenoviral vector were observed for HFB after 10 days and HaCaT after 30 days. Reporter gene expression in vivo was significantly higher in burned skin compared with unburned skin (P=0,004). Gene expression decreases from 2 to 7 days with no significant expression after 14 days. This study demonstrates that effective adenoviral-mediated gene transfer of epidermal primary cells and cell-lines is feasible. Ex vivo gene transfer in epithelial cells might have promise for the use in severely burned patients who receive autologous keratinocyte sheets. Transient cutaneous gene delivery in burn wounds using adenoviral vectors causes significant concentrations in the wound tissue for at least 1 week. Based on these findings, we hypothesize that transient cutaneous adenoviral gene delivery of wound healing promoting factors has potential for clinical application.

Adenoviridae↗