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Living donor liver transplantation was first developed to mitigate the limited access to deceased donor organs in Asia in the 1990s. This alternative liver transplantation method has become a widely practiced and established transplantation option for adult patients suffering with end-stage liver disease, and it has successfully helped address the shortage of deceased donors. The Society for the Advancement of Transplant Anesthesia and the Korean Society of Transplantation Anesthesiologists jointly reviewed published studies on the perioperative management of adult live liver donors undergoing donor hemi-hepatectomy. The goal of the review is to offer transplant anesthesiologists and critical care physicians a comprehensive overview of the perioperative management of adult live donors. We featured the current status, donor selection process, outcomes and complications, surgical procedure, anesthetic management, Enhanced Recovery After Surgery protocols, avoidance of blood transfusion, and considerations for emergency donation. Recent surgical advances, including laparoscopic donor hemi-hepatectomy and robotic laparoscopic donor surgery, are also addressed.
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Doença Hepática Terminal , Transplante de Fígado , Adulto , Doença Hepática Terminal/cirurgia , Hepatectomia/métodos , Humanos , Transplante de Fígado/métodos , Doadores Vivos , Coleta de Tecidos e ÓrgãosRESUMO
Efficient coupling to integrated high-quality-factor cavities is crucial for the employment of germanium quantum dot (QD) emitters in future monolithic silicon-based optoelectronic platforms. We report on strongly enhanced emission from single Ge QDs into L3 photonic crystal resonator (PCR) modes based on precise positioning of these dots at the maximum of the respective mode field energy density. Perfect site control of Ge QDs grown on prepatterned silicon-on-insulator substrates was exploited to fabricate in one processing run almost 300 PCRs containing single QDs in systematically varying positions within the cavities. Extensive photoluminescence studies on this cavity chip enable a direct evaluation of the position-dependent coupling efficiency between single dots and selected cavity modes. The experimental results demonstrate the great potential of the approach allowing CMOS-compatible parallel fabrication of arrays of spatially matched dot/cavity systems for group-IV-based data transfer or quantum optical systems in the telecom regime.
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Recently, it was shown that lasing from epitaxial Ge quantum dots (QDs) on Si substrates can be obtained if they are partially amorphized by Ge ion bombardment (GIB). Here, we present a model for the microscopic origin of the radiative transitions leading to enhanced photoluminescence (PL) from such GIB-QDs. We provide an energy level scheme for GIB-QDs in a crystalline Si matrix that is based on atomistic modeling with Monte Carlo (MC) analysis and density functional theory (DFT). The level scheme is consistent with a broad variety of PL experiments performed on as-grown and annealed GIB-QDs. Our results show that an extended point defect consisting of a split-[110] self-interstitial surrounded by a distorted crystal lattice of about 45 atoms leads to electronic states at the Γ-point of the Brillouin zone well below the conduction band minimum of crystalline Ge. Such defects in Ge QDs allow direct transitions of electrons localized at the split-interstitial with holes confined in the Ge QD. We identify the relevant growth and annealing parameters that will let GIB-QDs be employed as an efficient laser active medium.
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BACKGROUND: Wound infection can impair postoperative healing. Topical antibiotics have potential to treat wound infection and inflammation and minimize the adverse effects associated with systemic antibiotics. METHODS: Full-thickness porcine wounds were infected with Staphylococcus aureus. Using polyurethane wound enclosure devices, wounds were treated with topical 100 µg/ml minocycline, topical 1000 µg/ml minocycline, topical saline control, or 4 mg/kg intravenous minocycline. Bacteria were quantified in wound tissue and fluid obtained over 9 hours. Immunosorbent assays were used to analyze inflammatory marker concentrations. Minocycline's effect on in vitro migration and proliferation of human keratinocytes and fibroblasts was tested using scratch assays and metabolic assays, respectively. RESULTS: After 6 hours, 100 and 1000 µg/ml topical minocycline decreased bacteria in wound tissue to 3.5 ± 0.87 and 2.9 ± 2.3 log colony-forming units/g respectively, compared to 8.3 ± 0.9 log colony-forming units/g in control wounds (p < 0.001) and 6.9 ± 0.2 log colony-forming units/g in wounds treated with 4 mg/kg intravenous minocycline (p < 0.01). After 2 hours, topical minocycline reduced concentrations of the inflammatory cytokines interleukin-1ß, interleukin-6, and tumor necrosis factor-α (p < 0.01), and inflammatory cell counts in wound tissue (p < 0.05). In noninfected wounds, topical minocycline significantly reduced interleukin-1ß, interleukin-6, and inflammatory cell counts after 4 hours (p < 0.01). Matrix metalloproteinase-9 concentrations decreased after 1-hour treatment (p < 0.05). Keratinocyte and fibroblast in vitro functions were not adversely affected by 10 µg/ml minocycline or less. CONCLUSIONS: Topical minocycline significantly reduces bacterial burden and inflammation in infected wounds compared with wounds treated with intravenous minocycline or control wounds. Minocycline also decreases local inflammation independently of its antimicrobial effect.
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Antibacterianos/administração & dosagem , Inflamação/tratamento farmacológico , Minociclina/administração & dosagem , Infecções Estafilocócicas/tratamento farmacológico , Infecção dos Ferimentos/tratamento farmacológico , Administração Tópica , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Biomarcadores/metabolismo , Feminino , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Inflamação/etiologia , Inflamação/metabolismo , Injeções Intravenosas , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Minociclina/farmacologia , Minociclina/uso terapêutico , Distribuição Aleatória , Infecções Estafilocócicas/complicações , Infecções Estafilocócicas/metabolismo , Suínos , Resultado do Tratamento , Infecção dos Ferimentos/complicações , Infecção dos Ferimentos/metabolismoRESUMO
Semiconductor light-emitters compatible with standard Si integration technology (SIT) are of particular interest for overcoming limitations in the operating speed of microelectronic devices. Light sources based on group IV elements would be SIT-compatible, but suffer from the poor optoelectronic properties of bulk Si and Ge. Here we demonstrate that epitaxially grown Ge quantum dots (QDs) in a defect-free Si matrix show extraordinary optical properties if partially amorphized by Ge-ion bombardment (GIB). In contrast to conventional SiGe nanostructures, these QDs exhibit dramatically shortened carrier lifetimes and negligible thermal quenching of the photoluminescence (PL) up to room temperature. Microdisk resonators with embedded GIB-QDs exhibit threshold behavior as well as a superlinear increase of the integrated PL intensity with concomitant line width narrowing as the pump power increases. These findings demonstrate light amplification by stimulated emission in a fully SIT-compatible group IV nanosystem.
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Burn and blast injuries are frequently complicated by invasive infections, which lead to poor wound healing, delay in treatment, disability, or death. Traditional approach centers on early debridement, fluid resuscitation, and adjunct intravenous antibiotics. These modalities often prove inadequate in burns, where compromised local vasculature limits the tissue penetration of systemic antibiotics. Here, we demonstrate the treatment of infected burns with topical delivery of ultrahigh concentrations of antibiotics. Standardized burns were inoculated with Staphylococcus aureus or Pseudomonas aeruginosa. After debridement, burns were treated with either gentamicin (2 mg/mL) or minocycline (1 mg/mL) at concentrations greater than 1,000 times the minimum inhibitory concentration. Amount of bacteria was quantified in tissue biopsies and wound fluid following treatment. After six days of gentamicin or minocycline treatment, S. aureus counts decreased from 4.2 to 0.31 and 0.72 log CFU/g in tissue, respectively. Similarly, P. aeruginosa counts decreased from 2.5 to 0.0 and 1.5 log CFU/g in tissue, respectively. Counts of both S. aureus and P. aeruginosa remained at a baseline of 0.0 log CFU/mL in wound fluid for both treatment groups. The findings here demonstrate that super-therapeutic concentrations of antibiotics delivered topically can rapidly reduce bacterial counts in infected full-thickness porcine burns. This treatment approach may aid wound bed preparation and accelerate time to grafting.
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Antibacterianos/administração & dosagem , Antibacterianos/farmacologia , Queimaduras/tratamento farmacológico , Queimaduras/microbiologia , Infecções por Pseudomonas/tratamento farmacológico , Infecções Estafilocócicas/tratamento farmacológico , Infecção dos Ferimentos/tratamento farmacológico , Administração Tópica , Animais , Queimaduras/patologia , Desbridamento , Modelos Animais de Doenças , Feminino , Gentamicinas/administração & dosagem , Gentamicinas/farmacologia , Minociclina/administração & dosagem , Minociclina/farmacologia , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/patologia , Pseudomonas aeruginosa/efeitos dos fármacos , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/patologia , Staphylococcus aureus/efeitos dos fármacos , Suínos , Cicatrização/efeitos dos fármacos , Infecção dos Ferimentos/microbiologia , Infecção dos Ferimentos/patologiaRESUMO
Cell migration requires spatiotemporal integration of signals that regulate cytoskeletal dynamics. In response to a migration-promoting agent, cells begin to polarise and extend protrusions in the direction of migration. These cytoskeletal rearrangements are orchestrated by a variety of proteins, including focal adhesion kinase (FAK) and the Rho family of GTPases. CCN2, also known as connective tissue growth factor, has emerged as a regulator of cell migration but the mechanism by which CCN2 regulates keratinocyte function is not well understood. In this article, we sought to elucidate the basic mechanism of CCN2-induced cell migration in human keratinocytes. Immunohistochemical staining was used to demonstrate that treatment with CCN2 induces a migratory phenotype through actin disassembly, spreading of lamellipodia and re-orientation of the Golgi. In vitro assays were used to show that CCN2-induced cell migration is dependent on FAK, RhoA and Cdc42, but independent of Rac1. CCN2-treated keratinocytes displayed increased Cdc42 activity and decreased RhoA activity up to 12 hours post-treatment, with upregulation of p190RhoGAP. An improved understanding of how CCN2 regulates cell migration may establish the foundation for future therapeutics in fibrotic and neoplastic diseases.
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Movimento Celular , Actinas , Polaridade Celular , Fator de Crescimento do Tecido Conjuntivo , Proteína-Tirosina Quinases de Adesão Focal , Fatores de Troca do Nucleotídeo Guanina , Humanos , Queratinócitos , Proteínas Repressoras , Proteína cdc42 de Ligação ao GTPRESUMO
BACKGROUND: Injury to the skin can predispose individuals to invasive infection. The standard of care for infected wounds is treatment with intravenous antibiotics. However, antibiotics delivered intravenously may have poor tissue penetration and be dose limited by systemic side effects. Topical delivery of antibiotics reduces systemic complications and delivers increased drug concentrations directly to the wound. METHODS: Porcine full-thickness wounds infected with Staphylococcus aureus were treated with ultrahigh concentrations (over 1000 times the minimum inhibitory concentration) of gentamicin using an incubator-like wound healing platform. The aim of the present study was to evaluate clearance of infection and reduction in inflammation following treatment. Gentamicin cytotoxicity was evaluated by in vitro assays. RESULTS: Application of 2000 µg/ml gentamicin decreased bacterial counts in wound tissue from 7.2 ± 0.3 log colony-forming units/g to 2.6 ± 0.6 log colony-forming units/g in 6 hours, with no reduction observed in saline controls (p < 0.005). Bacterial counts in wound fluid decreased from 5.7 ± 0.9 log colony-forming units/ml to 0.0 ± 0 log colony-forming units/ml in 1 hour, with no reduction observed in saline controls (p < 0.005). Levels of interleukin-1ß were significantly reduced in gentamicin-treated wounds compared with saline controls (p < 0.005). In vitro, keratinocyte migration and proliferation were reduced at gentamicin concentrations between 100 and 1000 µg/ml. CONCLUSIONS: Topical delivery of ultrahigh concentrations of gentamicin rapidly decontaminates acutely infected wounds and maintains safe systemic levels. Treatment of infected wounds using the proposed methodology protects the wound and establishes a favorable baseline for subsequent treatment.
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Antibacterianos/administração & dosagem , Gentamicinas/administração & dosagem , Infecções Estafilocócicas/tratamento farmacológico , Infecção dos Ferimentos/tratamento farmacológico , Administração Tópica , Animais , Feminino , SuínosRESUMO
Highly photoconductive thin films of inorganic-capped PbS nanocrystal quantum dots (QDs) are reported. Stable colloidal dispersions of (NH4)3AsS3-capped PbS QDs were processed by a conventional dip-coating technique into a thin homogeneous film of electronically coupled PbS QDs. Upon drying at 130 °C, (NH4)3AsS3 capping ligands were converted into a thin layer of As2S3, acting as an infrared-transparent semiconducting glue. Photodetectors obtained by depositing such films onto glass substrates with interdigitate electrode structures feature extremely high light responsivity and detectivity with values of more than 200 A/W and 1.2×10(13) Jones, respectively, at infrared wavelengths up to 1400 nm. Importantly, these devices were fabricated and tested under ambient atmosphere. Using a set of time-resolved optoelectronic experiments, the important role played by the carrier trap states, presumably localized on the arsenic-sulfide surface coating, has been elucidated. Foremost, these traps enable a very high photoconductive gain of at least 200. The trap state density as a function of energy has been plotted from the frequency dependence of the photoinduced absorption (PIA), whereas the distribution of lifetimes of these traps was recovered from PIA and photoconductivity (PC) phase spectra. These trap states also have an important impact on carrier dynamics, which led us to propose a kinetic model for trap state filling that consistently describes the experimental photoconductivity transients at various intensities of excitation light. This model also provides realistic values for the photoconductive gain and thus may serve as a useful tool to describe photoconductivity in nanocrystal-based solids.
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We previously demonstrated that intratracheally administered S-ketamine inhibits alveolar fluid clearance (AFC), whereas an intravenous (i.v.) bolus injection had no effect. The aim of the present study was to characterize whether continuous i.v. infusion of S-ketamine, yielding clinically relevant plasma concentrations, inhibits AFC and whether its effect is enhanced in acute lung injury (ALI) which might favor the appearance of i.v. S-ketamine at the alveolar surface. AFC was measured in fluid-instilled rat lungs. S-ketamine was administered i.v. over 6 h (loading dose: 20 mg/kg, followed by 20 mg/kg/h), or intratracheally by addition to the instillate (75 µg/ml). ALI was induced by i.v. lipopolysaccharide (LPS; 7 mg/kg). Interleukin (IL)-6 and cytokine-induced neutrophil chemoattractant (CINC)-3 were measured by ELISA in plasma and bronchoalveolar lavage fluid. Isolated rat alveolar type-II cells were exposed to S-ketamine (75 µg/ml) and/or LPS (1 mg/ml) for 6 h, and transepithelial ion transport was measured as short circuit current (ISC). AFC was 27±5% (mean±SD) over 60 min in control rats and was unaffected by i.v. S-ketamine. Tracheal S-ketamine reduced AFC to 18±9%. In LPS-treated rats, AFC decreased to 16±6%. This effect was not enhanced by i.v. S-ketamine. LPS increased IL-6 and CINC-3 in plasma and bronchoalveolar lavage fluid. In alveolar type-II cells, S-ketamine reduced ISC by 37% via a decrease in amiloride-inhibitable sodium transport. Continuous administration of i.v. S-ketamine does not affect rat AFC even in endotoxin-induced ALI. Tracheal application with direct exposure of alveolar epithelial cells to S-ketamine decreases AFC by inhibition of amiloride-inhibitable sodium transport.
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Ketamina/efeitos adversos , Alvéolos Pulmonares/patologia , Edema Pulmonar/patologia , Administração Intravenosa , Animais , Transporte Biológico/efeitos dos fármacos , Líquido da Lavagem Broncoalveolar , Ketamina/administração & dosagem , Ketamina/sangue , Masculino , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Sódio/metabolismoRESUMO
OBJECTIVE: In the United States, around 50% of all musculoskeletal injuries are soft tissue injuries including ligaments and tendons. The objective of this study is to assess the role of amnion-derived cellular cytokine solution (ACCS) in carboxy-methyl cellulose (CMC) gel in the healing of Achilles tendon in a rat model, and to examine its effects on mechanical properties and collagen content. METHODS: Achilles tendons of Sprague-Dawley rats were exposed and transected. The distal and proximal ends were injected with either saline or ACCS in CMC, in a standardized fashion, and then sutured using a Kessler technique. Tendons from both groups were collected at 1, 2, 4, 6, and 8 weeks postoperatively and assessed for material properties. Collagen studies were performed, including collagen content, collagen cross-linking, tendon hydration, and immunohistochemistry. Tendons were also evaluated histologically for cross-sectional area. RESULTS: Mechanical testing demonstrated that treatment with ACCS in CMC significantly enhances breaking strength, ultimate tensile strength, yield strength, and Young's modulus in the tendon repair at early time points. In context, collagen content, as well as collagen cross-linking, was also significantly affected by the treatment. CONCLUSION: The application of ACCS in CMC has a positive effect on healing tendons by improving mechanical properties at early time points. Previous studies on onetime application of ACCS (not in CMC) did not show significant improvement on tendon healing at any time point. Therefore, the delivery in a slow release media like CMC seems to be essential for the effects of ACCS demonstrated in this study.
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The process of wound healing is dynamic and takes place over months to years, during which there is a resolution of angiogenesis, continued wound contraction, and connective tissue remodeling. The outcome of this process is most commonly the formation of a scar, defined as a fibrous tissue replacing normal tissues destroyed by injury or disease. Scars often have a lowered or total loss of vital skin functions and imbue a large burden on both the patient and the health care system as a whole. Scar treatments are plentiful but are often unsatisfactory or inconsistent. No single treatment method has been universally adopted. To evaluate the clinical treatment as well as research focused on developing novel methods for scar management, objective studies of the progression of scar formation and the properties of mature scars are needed. Several parameters, including barrier function as well as mechanical and physiological properties, need to be taken into account when both categorizing and treating healing wounds and scars. To date, there is no available methodology that provides a comprehensive evaluation of a scar's properties. This review aims at presenting an overview of available scar assessment methods and devices, ranging from analysis of collagen properties in tissue biopsies to noninvasive methods for studies of mechanical parameters such as breaking strength and skin elasticity. In the cases where conclusive studies have been performed, the differences between normal skin and scar with respect to the above parameters are presented. Furthermore, this review highlights areas where the development of additional modalities are needed.
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Cicatriz/fisiopatologia , Matriz Extracelular/ultraestrutura , Pele/fisiopatologia , Cicatrização , Cicatriz/patologia , Cicatriz/prevenção & controle , Colágeno/metabolismo , Elasticidade , Humanos , Pele/lesões , Fenômenos Fisiológicos da Pele , Transplante de Pele/métodos , Pele Artificial , Resultado do TratamentoRESUMO
Transplantation of skin micrografts in a 1:100 ratio regenerate the epidermis of full-thickness wounds in pigs within 14 days in a wet environment. The aim of the current study was to combine micrografts and commercially available moist dressings. We hypothesized that micrografts regenerate the epidermis when covered with a moist dressing. 5cm×5cm and 10cm×10cm full-thickness wounds were created on the backs of pigs. Wounds were transplanted with 0.8mm×0.8mm micrografts created from a split-thickness skin graft in a 1:100 ratio. 5cm×5cm wounds were treated with wound chambers, moist dressings or dry gauze (non-transplanted control group). 10cm×10cm wounds were compared to non-transplanted wounds, both covered with moist dressings. Reepithelialization was assessed in biopsies from day 10, 14 and 18 post-transplantation. 5cm×5cm transplanted wounds covered with moist dressings showed 69.5±20.6% reepithelialization by day 14 and 90.5±10.4% by day 18, similar to wounds covered with a wound chamber (63.9±16.7 and 86.2±11.9%, respectively). 18 days post-transplantation, 10cm×10cm transplanted wounds covered with moist dressings showed 66.1±10.3% reepithelialization, whereas nontransplanted wounds covered with moist dressings were 40.6±6.6% reepithelialized. We conclude that micrografts combined with clinically available moist dressings regenerate the epidermis of full-thickness wounds.
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Curativos Hidrocoloides , Reepitelização , Transplante de Pele/métodos , Pele/lesões , Animais , Epiderme/patologia , Feminino , Suínos , Resultado do Tratamento , Técnicas de Fechamento de Ferimentos , CicatrizaçãoRESUMO
BACKGROUND: CCN2 (previously known as connective tissue growth factor) is a multifunctional matricellular protein that has numerous effects on cell life and cell interactions with the connective tissue. Although the importance of CCN2 for the fibrotic process in wound healing has been well studied, the involvement of CCN2 in keratinocyte function has not yet been explored. Therefore, the aim of the present study was to investigate the role of CCN2 in the epidermis during wound healing. MATERIALS AND METHODS: Immunohistochemistry was done on sections from full-thickness porcine wounds. The effect of CCN2 on the migration of cultured human keratinocytes exposed to scratch wounds, the effect on phosphorylation of extracellular signal-related kinases (ERK), and the effect of adding inhibitors to the ERK/mitogen-activated protein kinase pathway to human keratinocytes were studied. RESULTS: The CCN2 protein was transiently expressed in vivo at the leading keratinocyte edge during re-epithelialization of full-thickness porcine wounds. In vitro, exogenous addition of CCN2 to human keratinocyte cultures regulated keratinocyte migration and resulted in phosphorylation of ERK. The addition of inhibitors of ERK/mitogen-activated protein kinase counteracted the effect of CCN2 on migration. CONCLUSIONS: CCN2 was transiently expressed at the leading keratinocyte edge in vivo. The biologic importance of this was supported in vitro, because CCN2 regulated human keratinocyte migration through activation of the Ras-mitogen-activated protein kinase kinase-ERK signal transduction pathway.
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Movimento Celular/fisiologia , Fator de Crescimento do Tecido Conjuntivo/metabolismo , Queratinócitos/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Cicatrização/fisiologia , Ferimentos e Lesões/metabolismo , Animais , Proliferação de Células , Células Cultivadas , Células Epidérmicas , Epiderme/lesões , Epiderme/metabolismo , Feminino , Imunofluorescência , Humanos , Técnicas In Vitro , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Sus scrofa , Ferimentos e Lesões/patologia , Proteínas ras/metabolismoRESUMO
OBJECTIVE: Tendon injuries produce considerable morbidity, long-lasting disability, and remain a considerable challenge for clinicians and patients. The objective of the study was to assess the effect of amnion-derived multipotent progenitor (AMP) cells and amnion-derived cell cytokine solution on Achilles tendon healing by using a rat model. METHODS: Achilles tendons of Sprague-Dawley rats were exposed and transected. The distal and proximal ends were injected with either saline, amnion-derived cell cytokine solution, or AMP cells in a standardized fashion and then sutured by using a Kessler technique. Tendons from each group (n = 6-13) were collected at weeks 1, 2, and 4 postoperatively and assessed for material properties (ultimate tensile strength, Young modulus, yield strength, and breaking strength). Tendons were also evaluated histologically for cross-sectional area by using hematoxylin-eosin and trichrome stains. RESULTS: Mechanical testing showed that the Young modulus was significantly higher in AMP cells-treated tendons at week 4 compared with both saline-treated and amnion-derived cell cytokine solution-treated tendons. Yield strength was significantly higher in the AMP cells-treated group compared with saline-treated controls at week 4. No significant differences were observed between the study groups at weeks 1 and 2. DISCUSSION: Amnion-derived multipotent progenitor cells have a positive effect on healing tendons by improving mechanical strength and elastic modulus during the healing process. The presented findings suggest the clinical utility of AMP cells in facilitating the healing of ruptured tendons. Both the Young modulus and yield strengths of tendons increased significantly following treatment with AMP cells.
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BACKGROUND: Major loss of skin following burns or trauma requires skin grafting for repair. In addition, chronic wounds frequently require skin grafts. Current treatments are either cumbersome, limited in possible expansion ratio, costly, or require extensive time for treatment. This study investigates a new way of regenerating skin after major burns and other trauma, providing 100-fold expansion of a split-thickness skin graft. METHODS: Submillimeter micrografts were created by controlled mincing of a split-thickness skin graft and transplanted to porcine full-thickness wounds. By creating an incubator-like microenvironment using wound chambers, the micrografts provide reepithelialization whether placed dermal side up or dermal side down. RESULTS: Transplantation of micrografts in a 1:100 expansion ratio results in complete epithelialization of both healthy and diabetic wounds within 14 days. In comparison, nontransplanted wounds showed 62 percent reepithelialization in healthy pigs and 49 percent in diabetic pigs at the corresponding time point. CONCLUSIONS: Minced skin micrografts are very effective in wound repair and can provide 100-fold expansion of a skin graft. Early clinical results confirm the utility of this technique.
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Epiderme/fisiologia , Microcirurgia , Regeneração , Transplante de Pele/métodos , Animais , Feminino , Suínos , CicatrizaçãoRESUMO
BACKGROUND: Transplantation of skin micrografts (MGs), split-thickness skin grafts (STSGs), or cultured autologous keratinocytes (CKs) enhances the healing of large full-thickness wounds. This study compares these methods in a porcine wound model, investigating the utility of micrograft transplantation in skin restoration. STUDY DESIGN: Full-thickness wounds were created on Yorkshire pigs and assigned to one of the following treatment groups: MGs, STSGs, CKs, wet nontransplanted, or dry nontransplanted. Dry wounds were covered with gauze and the other groups' wounds were enclosed in a polyurethane chamber containing saline. Biopsies were collected 6, 12, and 18 days after wounding. Quantitative and qualitative wound healing parameters including macroscopic scar appearance, wound contraction, neoepidermal maturation, rete ridge formation, granulation tissue thickness and width, and scar tissue formation were studied. RESULTS: Transplanted wounds scored lower on the Vancouver Scar Scale compared with nontransplanted wounds, indicating a better healing outcome. All transplanted wounds exhibited significantly lower contraction compared with nontransplanted wounds. Wounds transplanted with either MGs, STSGs, or CKs showed a significant increase in re-epithelialization compared with nontransplanted wounds. Wounds transplanted with MGs or STSGs exhibited improved epidermal healing compared with nongrafted wounds. Furthermore, transplantation with STSGs or MGs led to less scar tissue formation compared with the nontransplanted wounds. No significant impact on scar formation was observed after transplantation of CKs. CONCLUSIONS: Qualitative and quantitative measurements collected from full-thickness porcine wounds show that transplantation of MGs improve wound healing parameters and is comparable to treatment with STSGs.
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Queratinócitos/transplante , Transplante de Pele , Cicatrização/fisiologia , Ferimentos Penetrantes/terapia , Animais , Transplante de Células , Células Cultivadas , Modelos Animais de Doenças , Feminino , Suínos , Fatores de Tempo , Transplante Autólogo , Ferimentos Penetrantes/patologiaRESUMO
We show that both the morphology and the optoelectronic properties of SiGe islands growing in the pits of periodically pre-patterned Si(001) substrates are determined by the amount of Ge deposited per unit cell of the pattern. Pit-periods (p) ranging from 300 to 900 nm were investigated, and Ge growth was performed by molecular beam epitaxy (MBE) at temperatures of 690 and 760 °C. The ordered SiGe islands show photoluminescence (PL) emission, which becomes almost completely quenched, once a critical island volume is exceeded. By atomic force and transmission electron microscope images we identify the transition from pyramid-shaped to dome-shaped islands with increasing p. Eventually, the nucleation of dislocations in the islands leads to PL quenching. Below a critical Ge coverage a narrowing and a blue shift of the PL emission is observed, as compared to islands grown on a planar reference area of the same sample.
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Germânio/química , Medições Luminescentes/métodos , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Silício/química , Teste de Materiais , Tamanho da PartículaRESUMO
BACKGROUND: Transplantation of genetically modified keratinocytes has been shown to accelerate wound healing. However, this method is labor-intensive and time-consuming. We have developed a new technique of intraoperative gene delivery to wounds that involves transplantation of transfected minced skin particles (MSPs) derived from harvested partial-thickness skin. STUDY DESIGN: MSPs measuring 0.8 × 0.8 × 0.35 mm were created from a split-thickness skin graft of a pig. In vitro transfection was carried out with adenoviral LacZ (Ad-LacZ) for qualitative and adenoviral vascular endothelial growth factor (Ad-VEGF) for quantitative analysis. Transfected MSPs were transplanted to each of 2.5 × 2.5 cm full-thickness wounds on the dorsum of the pig. Nontransfected MSPs served as controls. Wound chambers were applied and injected with saline to create a wet environment. RESULTS: LacZ expression was detected in migrating cells originating from MSPs both in vitro and in vivo. VEGF expression in the wound fluid of Ad-VEGF-MSP-transplanted wounds on each of days 2 to 4 (mean ± SEM 6.74 ± 1.89 ng/mL, day 2; 9.88 ± 2.27 ng/mL, day 3; 9.87 ± 1.28 ng/mL, day 4) was significantly higher (p < 0.0001) compared with wounds transplanted with either untransfected MSPs, Ad-LacZ-MSPs, or untransplanted controls. In vitro VEGF expression was significantly higher (p < 0.0001) in Ad-VEGF 1 × 10(10) transfected MSPs compared with either Ad-VEGF 1 × 10(9) transfected MSPs or untransfected MSPs. Wounds transplanted with Ad-VEGF-MSPs showed significantly higher (p < 0.0001) numbers of newly formed blood vessels (12.6 ± 0.9 vessels/high power field [HPF]) compared with wounds transplanted with either Ad-LacZ-MSPs (4.4 ± 0.5 vessels/HPF) or untransfected MSPs (5.2 ± 0.7 vessels/HPF). All MSP-transplanted wounds (Ad-VEGF-MSPs, untransfected MSPs, Ad-LacZ-MSPs) showed significantly higher re-epithelialization compared with untransplanted wounds on days 10 and 14 (p < 0.0001). CONCLUSIONS: We demonstrated successful transfection of MSPs that can be transplanted to wounds as a source of gene-expressing cells. This technique can be used to deliver growth-modulating genes in wound healing.
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Técnicas de Transferência de Genes , Neovascularização Fisiológica/genética , Fator A de Crescimento do Endotélio Vascular/genética , Cicatrização/genética , Animais , Modelos Animais de Doenças , Feminino , Transplante de Pele , SuínosRESUMO
High yields of pure and viable porcine islet cells (PICs) to be used for microencapsulation are crucial for successful xenotransplantation. Mechanical disruption of the pancreas, enzymes used for digestion, digestion temperature and time are among the factors known to cause oxidative stress and to impact on the yield, purity and viability of PICs. The aim of our study was to optimize conventional procedures in order to minimize the oxidative stress that occurs during the isolation and purification of PICs. Porcine pancreatic tissue was harvested at a local slaughterhouse, and 15 consecutive isolations of PICs were performed with a modified automated Ricordi method (Graz method) using a shorter digestion time, a lower digestion temperature and minimal mechanical stress. PICs were purified with the Lymphoprep density gradient medium. Purity and viability were assessed immediately after the isolation process and after overnight culture. PIC function was tested in glucose stimulation experiments and insulin concentration was determined by ELISA. Oxidative stress was assessed by measuring isoprostanes (IP), malondialdehyde (MDA) and lipase levels using a HPLC-based, colorimetric liquid assay or ELISA, respectively. The mean yield of PICs was 3479 +/- 542 IEQs/g pancreas, with 96.4% viability and 97.7% purity. There was no significant loss in PIC viability after overnight culture. Insulin secretion in response to glucose was not impaired after isolation and purification. IP, MDA and lipase levels did not change significantly during the isolation procedure. With our new Graz method we seem to have succeeded in preventing oxidative stress and achieving high yields of pure and viable PICs.