scholarly journals Dermal fibroblast-like cells reprogrammed directly from adipocytes in mouse

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Mitsunobu Toyosaki ◽  
Koichiro Homma ◽  
Sayuri Suzuki ◽  
Naoto Muraoka ◽  
Hisayuki Hashimoto ◽  
...  

AbstractIn deep burns, early wound closure is important for healing, and skin grafting is mainly used for wound closure. However, it is difficult to achieve early wound closure in extensive total body surface area deep burns due to the lack of donor sites. Dermal fibroblasts, responsible for dermis formation, may be lost in deep burns. However, fat layers composed of adipocytes, lying underneath the dermis, are retained even in such cases. Direct reprogramming is a novel method for directly reprograming some cells into other types by introducing specific master regulators; it has exhibited appreciable success in various fields. In this study, we aimed to assess whether the transfection of master regulators (ELF4, FOXC2, FOXO1, IRF1, PRRX1, and ZEB1) could reprogram mouse adipocytes into dermal fibroblast-like cells. Our results indicated the shrinkage of fat droplets in reprogrammed mouse adipocytes and their transformation into spindle-shaped dermal fibroblasts. Reduced expression of PPAR-2, c/EBP, aP2, and leptin, the known markers of adipocytes, in RT-PCR, and enhanced expression of anti-ER-TR7, the known anti-fibroblast marker, in immunocytochemistry, were confirmed in the reprogrammed mouse adipocytes. The dermal fibroblast-like cells, reported here, may open up a new treatment mode for enabling early closure of deep burn wounds.

2021 ◽  
Vol 42 (Supplement_1) ◽  
pp. S184-S185
Author(s):  
Eric Curfman ◽  
Anjay Khandelwal

Abstract Introduction Calcific Uremic Arteriolopathy (CUA), commonly known as Calciphylaxis, is a rare disorder characterized by ischemic necrosis of the skin and histologically by arteriolar calcification. CUA is most commonly seen in patients with end-stage renal disease (ESRD) but can be seen in other patients as well. CUA carries an extremely high mortality rate, with up to 80% in some studies, even in patients with limited disease. In light of this, many surgeons have adopted a “do-not-touch” practice with these patients. Over the past several years, our institution has seen an increase in referrals for the management of large total-body-surface-area (TBSA) CUA. Methods Retrospective review of all patients with biopsy-proven (by dermatopathology) large TBSA (>=5% TBSA) CUA admitted to a Verified Adult and Pediatric Burn Center from 2015 to present. Demographics, laboratory data, treatment modalities and outcomes including mortality and wound closure were recorded. Results A total of 8 patients with large TBSA CUA were admitted after being transferred from outside hospitals. Average TBSA affected was 13.76% (SD 7.27). 6 of these patients (75%) were noted to have non-uremic calciphylaxis. All patients had positive wound cultures on admission, and 1 patient (12.5%) developed a bacteremia in hospital. There were no central line associated bloodstream infections, catheter associated urinary tract infections or ventilator associated infections. All patients underwent surgical debridement (average 4.125, range 2–5), and 5 patients (62.5%) underwent grafting, (average 1.6, range 2–5) and subsequently proceeded to wound closure. In-hospital mortality was 25% and another patient was referred to a hospice facility after being readmitted with medical complications of her calciphylaxis. Secondary findings included 50% of the patients recently experienced significant weight loss (>100 lbs). On admission, 2 patients (25%) had abnormal serum calcium, 3 patients (37.5%) had abnormal serum PO4, and 4 (50%) patients had abnormal PTH levels. 2 patients (25%) had a recent exposure to warfarin (within 6 months). Conclusions Utilizing a multi-modal management strategy that includes surgical debridement and skin grafting, patients with calciphylaxis can progress to wound closure.


Author(s):  
Frank Faltraco ◽  
Denise Palm ◽  
Adriana Uzoni ◽  
Lena Borchert ◽  
Frederick Simon ◽  
...  

AbstractA link between dopamine levels, circadian gene expression, and attention deficit hyperactivity disorder (ADHD) has already been demonstrated. The aim of this study was to investigate the extent of these relationships by measuring circadian gene expression in primary human-derived dermal fibroblast cultures (HDF) after dopamine exposure. We analyzed circadian preference, behavioral circadian and sleep parameters as well as the circadian gene expression in a cohort of healthy controls and participants with ADHD. Circadian preference was evaluated with German Morningness-Eveningness-Questionnaire (D-MEQ) and rhythms of sleep/wake behavior were assessed via actigraphy. After ex vivo exposure to different dopamine concentrations in human dermal fibroblast (HDF) cultures, the rhythmicity of circadian gene expression (Clock, Bmal1, Per1-3, Cry1) was analyzed via qRT-PCR. We found no statistical significant effect in the actigraphy of both groups (healthy controls, ADHD group) for mid-sleep on weekend days, mid-sleep on weekdays, social jetlag, wake after sleep onset, and total number of wake bouts. D-MEQ scores indicated that healthy controls had no evening preference, whereas subjects with ADHD displayed both definitive and moderate evening preferences. Dopamine has no effect on Per3 expression in healthy controls, but produces a significant difference in the ADHD group at ZT24 and ZT28. In the ADHD group, incubation with dopamine, either 1 µM or 10 µM, resulted in an adjustment of Per3 expression to control levels. A similar effect also was found in the expression of Per2. Statistical significant differences in the expression of Per2 (ZT4) in the control group compared to the ADHD group were found, following incubation with dopamine. The present study illustrates that dopamine impacts on circadian function. The results lead to the suggestion that dopamine may improve the sleep quality as well as ADHD symptoms by adjustment of the circadian gene expression, especially for Per2 and Per3.


Processes ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 801
Author(s):  
Jehun Choi ◽  
Gwi-Yeong Jang ◽  
Jeonghoon Lee ◽  
Hae-Young Chung ◽  
Hyung-Jun Noh ◽  
...  

Senescence is the phenomenon by which physiological functions of organisms degenerate with time. Cellular senescence is marked by an inhibition of cell cycle progression. Beta-galactosidase accumulates in the lysosomes of aged cells. In this study, human dermal fibroblast cells (HDFs) were treated with 0.5 μM doxorubicin for 4 h to induce cellular senescence. Senescence-associated beta-galactosidase (SA-β-gal) activity was then measured 72 h after treatment with aerial parts of Dendranthema zawadskii var. lucidum (Nakai) J.H. Park (DZ) extract. Treatment with DZ extract significantly decreased SA-β-gal activity in a dose-dependent manner in HDFs. Additionally, DZ extract treatment reduced age-related oxidative stress and inflammation in the aortas of aged rats. The reactive oxygen species (ROS) levels in aortas of aged control rats were higher than those in young rats. However, DZ extract-fed aged rats showed significantly lower ROS levels than the aged control rats. When the aged rats were treated with DZ extract at either 0.2 or 1.0 mg∙kg−1∙day−1, NF-κB levels in aorta tissue decreased significantly compared to those in aorta tissue of the aged control rats without DZ treatment. In addition, DZ extract-fed aged rat aortas showed significant reductions in expression of iNOS and COX-2 induced by NF-κB translocation. Therefore, these results suggest that DZ effectively inhibited senescence-related NF-κB activation and inflammation. DZ extract may have a role in the prevention of the vascular inflammatory responses that occur during vascular aging.


Author(s):  
Praveen Kumar Arumugam ◽  
Vamseedharan Muthukumar ◽  
Rahul Bamal

Abstract Fasciotomy is indicated to relieve compartment syndrome caused by electric burns. Many techniques are available to close the fasciotomy wounds including vacuum-assisted closure, skin grafting, and healing by secondary intention. This study assessed the shoelace technique in fasciotomy wound closure in patients with electric burns. The study included 19 fasciotomy wounds that were treated by shoelace technique (Group ST, n = 10 fasciotomy wounds) or by skin grafting/healing by secondary intention (Group C, n = 9 fasciotomy wounds). Data were collected for wound surface area, time to intervention, time to wound closure, rate of decrease in wound surface area after application of shoelace technique and associated complications. The mean time to intervention after fasciotomy was significantly lower in Group ST—7.6 ± 3.8 days as compared to 15.8 ± 5.3 days in Group C (P = .004). The median time to closure was also significantly lower in Group ST—7 days (range 6–10) as compared to Group C—20 days (range 12–48) (P < .001). Primary closure was achieved in 80% cases in the group ST and no complications were recorded. The shoelace technique is an economical, fast, and effective method of fasciotomy wound closure in electric burns, especially in high volume centers and resource-limited areas.


2021 ◽  
Vol 35 ◽  
pp. 205873842110167
Author(s):  
Zhensen Zhu ◽  
Bo Chen ◽  
Liang Peng ◽  
Songying Gao ◽  
Jingdong Guo ◽  
...  

Activated M2 macrophages are involved in hypertrophic scar (HS) formation via manipulating the differentiation of fibroblasts to myofibroblasts having the proliferative capacity and biological function. However, the function of exosomes derived from M2 macrophages in HS formation is unclear. Thus, this study aims to investigate the role of exosomes derived by M2 in the formation of HS. To understand the effect of exosomes derived from M2 macrophages on formation of HS, M2 macrophages were co-cultured with human dermal fibroblast (HDF) cells. Cell Counting Kit-8 assay was performed to evaluate HDF proliferation. To evaluate the migration and invasion of HDFs, wound-healing and transwell invasion assays were performed, respectively. To investigate the interaction between LINC01605 and miR-493-3p, a dual-luciferase reporter gene assay was adopted; consequently, an interaction between miR-493-3p and AKT1 was detected. Our results demonstrated that exosomes derived from M2 macrophages promoted the proliferation, migration, and invasion of HDFs. Additionally, we found that long noncoding RNA LINC01605, enriched in exosomes derived from M2 macrophages, promoted fibrosis of HDFs and that GW4869, an inhibitor of exosomes, could revert this effect. Mechanistically, LINC01605 promoted fibrosis of HDFs by directly inhibiting the secretion of miR-493-3p, and miR-493-3p down-regulated the expression of AKT1. Exosomes derived from M2 macrophages promote the proliferation and migration of HDFs by transmitting LINC01605, which may activate the AKT signaling pathway by sponging miR-493-3p. Our results provide a novel approach and basis for further investigation of the function of M2 macrophages in HS formation.


Author(s):  
Xingxin Gao ◽  
Min Zhang ◽  
Yuan Lin ◽  
Dehui Li ◽  
Liming Zhang

Abstract Auto-skin grafting is the current treatment of choice for extensive burns. Nevertheless, the lack of donor sites for skin grafting remains one of the greatest limiting factors for the treatment of extensively burned patients. We present the case of a 53-year-old male patient with deep and full thickness burns on 91% of the total body surface area. We used the Meek technique for split-thickness skin graft expansion to treat this patient. In order to obtain sufficient skin for grafting, we repeatedly harvested the same anatomical areas. Acceleration of burn wounds, recipient, and donor site healing was achieved by systemic treatment with recombinant human growth hormone and topical recombinant human epidermal growth factors. This combined, complex treatment modality contributed to the successful skin repair in this patient.


2018 ◽  
Vol 12 (5) ◽  
pp. 480-487 ◽  
Author(s):  
K. B. L. Lim ◽  
T. Laine ◽  
J. Y. Chooi ◽  
W. K. Lye ◽  
B. J. Y. Lee ◽  
...  

Purpose Acute compartment syndrome (ACS) requires urgent fasciotomy to decompress the relevant muscle compartment/s prior to onset of irreversible myonecrosis and nerve injury. A fasciotomy is not a benign procedure. This study aims to describe and quantify early morbidity directly associated with fasciotomies for ACS in children. Methods Clinical charts of 104 children who underwent 112 fasciotomies over a 13-year period at a tertiary children’s hospital were reviewed. The following were analyzed: ACS aetiology, fasciotomy site, number of subsequent procedures, method of wound closure, short-term complications and length of hospital stay. Results Short-term complications included wound infections (6.7%) and the need for blood transfusion (7.7%). Median number of additional operations for wound closure was two (0 to 10) and median inpatient stay was 12 days (3 to 63; SD 11.7). After three unsuccessful attempts at primary closure, likelihood of needing skin grafting for coverage exceeded 80%. Analyses showed that fasciotomy-wound infections were associated with higher risk for four or more closure procedures. Number of procedures required for wound closure correlated with longer inpatient stay as did ACS associated with non-orthopaedic causes. Conclusion Fasciotomy is associated with significant early morbidity, the need for multiple closure operations, and prolonged hospital stay. The decision for fasciotomy needs careful consideration to avoid unnecessary fasciotomies, without increasing the risk of permanent injury from missed or delayed diagnosis. Skin grafting should be considered after three unsuccessful closure attempts. Less invasive tests or continuous monitoring (for high-risk patients) for compartment syndrome may help reduce unnecessary fasciotomies. Level of Evidence Level IV, Case series


NEMESIS ◽  
2018 ◽  
Vol 3 (1) ◽  
pp. 15 ◽  
Author(s):  
Marcin Kozakiewicz ◽  
Piotr Szymor ◽  
Raphael Olszewski

Objective: Our study aimed to determine the possibility of using models created with a low-cost, paper based 3D printer in an operating room. Therefore influence of different methods of sterilization on models was tested and cytotoxicity of generated models was determined. Material and methods: 30 cuboids divided into three groups were used for verification of shape stability after sterilization. Each group was sterilized either with: Ethylene oxide in temperature 55˚C, Hydrogen peroxide gas plasma in temperature 60˚C or Gamma irradiation at 21˚C, 25kGy. Each cuboid was measured using calliper three times before and three times after sterilization. Results were analysed statistically in Statgraphics Plus. Statistical significance was determined as p< 0.05. Sixty cylinders divided into six groups were used for cytotoxicity tests. Three of those groups were covered before sterilization with 2-octyl-cyanoacrylate. Each group was sterilized with one of the previously described methods. Cytotoxicity was tested by Nanostructural and Molecular Biophysics Laboratory in Technopark Lodz using normal adult human dermal fibroblasts. Survival of cells was tested using spectrophotometry with XTT and was defined as ratio of absorbency of tested probe to absorbency of control probe. Calcein/Ethidium dyeing test was performed according to LIVE/DEAD Viability/Cytotoxicity Kit protocol. Observation was done under Olympus GX71 fluorescence microscope. Results: There was no statistically significant difference for established statistical significance p=0.05 in cuboids dimensions before and after sterilization regardless of sterilization method. In XTT analysis all samples showed higher cytotoxicity against normal, human, adult dermal fibroblast culture when compared to positive control. ANOVA statistical analysis confirmed that 2-octyl cyanoacrylate coating of paper model improved biological behaviour of the material. It decreased cytotoxicity of the model independently of sterilization method. In calcein/ethidium dyeing test due to the high fluorescence of the background caused by cylinders of analysed substance it was impossible to perform the exact analysis of the number of marked cells. Conclusions: Acquired results allow to conclude that Mcor Technology Matrix 300 3D paper-based models can be used in operating room only if covered with cyanoacrylate tissue adhesive. Nemesis relevance: We found no statistically significant difference in cuboids dimensions before and after sterilization regardless of sterilization method. Three-dimensional paper-based models present with high cytotoxicity without coating.


2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Jutamas Somchaichana ◽  
Tanom Bunaprasert ◽  
Suthiluk Patumraj

Acanthus ebracteatusVahl. is a Thai herb that is effective in wound healing. We sought to quantitatively determine whether or not the combined application ofAcanthus ebracteatusVahl. and a collagen scaffold will increase wound closure and angiogenesis. Balb/c mice (body weight: 22–25 g) were anesthetized with sodium thiopental. The dorsal skin incision measuring 1.5 × 1.5 cm was made and then deepened using scissors to produce a full-thickness incision down to the level of the panniculus carnosus. The size of the wound was approximately 10% of the total body surface area. The collagen sheet was implanted onto the wound. Animals were divided into 4 major groups as follows: wound with normal saline (W-NSS), wound treated with 0.3 g/kg BW ofAcanthus ebracteatusVahl. extract (W-AE (0.3 g/kg.bw)), wound implanted with collagen scaffold (W-Coll), and wound implanted with collagen scaffold and treated with 0.3 g/kg BW ofAcanthus ebracteatusVahl. (W-Coll-AE combination). On day 14, the W-Coll-AE group showed decreased wound areas and increased capillary vascularity (CV) when compared to the other 3 groups, W-NSS, W-AE0.3, and W-Coll. In the present study, the combination of AE0.3 with collagen showed the best effect on skin angiogenesis and promoted wound closure with less neutrophil infiltration.


Sign in / Sign up

Export Citation Format

Share Document