BACKGROUND: Decellularized extracellular matrix (dECM) is a non-cellular scaffold with various functions in tissue engineering and regenerative medicine. Elastin is related to tissue elasticity and scarless wound healing, abundantly found in lung and blood vessel tissues. We studied the characteristics of blood vessel-derived dECM (VdECM) and its effect in wound healing. METHODS: VdECM was prepared from porcine blood vessel tissue. Weight percentages of elastin of VdECM and atelocollagen were analyzed. Migratory potential of VdECM was tested by scratch assay. VdECM in hydrogel form was microscopically examined, tested for fibroblast proliferation, and examined for L/D staining. Cytokine array of various growth factors in adipocyte-derived mesenchymal stem cell (ASC) media with VdECM was done. Animal wound model showed the wound healing effect of VdECM hydrogel in comparison to other topical agents. RESULTS: VdECM contained 6.7 times more elastin than atelocollagen per unit weight. Microscopic view of 0.35% VdECM hydrogel showed consistent distribution. Compared to 3% atelocollagen, 0.35% VdECM showed superior results in fibroblast migration. Fluorescent microscopic findings of L/D assay had highest percentage of cell survival in 1% VdECM compared to atelocollagen. Growth factor expression was drastically amplified when VdECM was added to ASC media. In the animal study model, epithelialization rate in the VdECM group was higher than that of control, oxytetracycline, and epidermal growth factor ointments. CONCLUSION VdECM contains a high ratio of elastin to collagen and amplifies expressions of many growth factors. It promotes fibroblast migration, proliferation, and survival, and epithelialization comparable to other topical agents.

BACKGROUND: The repair of large bone defects remains a significant challenge in clinical practice and requires bone grafts or substitute materials. In this study, we developed a unique hybrid bone scaffold comprising a three dimensional (3D)-printed metal plate for weight bearing and a biodegradable polymer tube serving as bone conduit. We assessed the long-term effect of the hybrid bone scaffold in repairing radial bone defects in a beagle model. METHODS: Bone defects were created surgically on the radial bone of three beagle dogs and individually-tailored scaffolds were used for reconstruction with or without injection of autologous bone and decellularized extracellular matrix (dECM). The repaired tissue was evaluated by X-ray, micro-computed tomography, and histological observation 6 months after surgery. The functional integrity of hybrid bone scaffold-mediated reconstructions was assessed by gait analysis. RESULTS: In vivo analysis showed that the hybrid bone scaffolds maintained the physical space and bone conductivity around the defect. New bone was formed adjacent to the scaffolds. Addition of autologous bone and dECM in the polymer tube improved healing by enhancing bone induction and osteoconduction. Furthermore, the beagles’ gait appeared normal by 4 months. CONCLUSION The future of bone healing and regeneration is closely related to advances in tissue engineering. Bone production using autologous bone and dECM loaded on 3D-printed hybrid bone scaffolds can successfully induce osteogenesis and provide mechanical force for functional bone regeneration, even in large bone defects.

Background: The stromal vascular fraction (SVF) isolated from adipose tissue, which contains stem cells as well as other cell types, has been applied in various research fields. Although different enzymatic concentrations and treatment durations have been applied to isolate the SVF, optimal conditions have not been established. Thus, we aimed to establish the optimal conditions for isolation of the SVF from adipose tissue by automated systems. Methods: The SVF was collected from removed adipose tissues of five donors during surgery. The SVF was treated with 0.1% or 0.2% collagenase type I for 20, 40, or 60 min. Then, colony forming unit (CFU) assays and flow cytometry were performed to characterize the adipose stem cells (ASCs). A cytokine array was used to investigate the correlation between colony-formation ability and the secretion of isolated ASCs. Results: Treatment with 0.1% collagenase type I for 60 min resulted in a higher SVF yield, whereas treatment with 0.1% collagenase for 40 min resulted in higher CFU values. In addition, expression of interleukin (IL)-6, IL-8, and monocyte chemoattractant protein-1 in the SVF was higher in the high-CFU group than in the low-CFU group. Conclusion The optimal conditions for isolation of the SVF from adipose tissue were treatment with 0.1% collagenase type I for 40 min. We identified the conditions required for efficient SVF isolation based on high CFU values, and our results will facilitate the development of automated systems.

This study was to investigate the effect of subcutaneous injection of the adipose stem cells (ASCs) with conditioned media (CM) in the treatment of acne vulgaris scar. We used Adult male New Zealand white rabbit ears as an animal model and induced acne formation by Kignman method. Adipose tissue was isolated and harvested from the scapula of rabbits, and ASCs were cultured and expanded until passage 1. There have four groups in our experiment, include phosphate buffered saline (PBS), ASCs with PBS (ASC + PBS), CM, and ASCs with CM (ASC + CM) group. This solution of 0.6 ml injected to subcutaneous in each group. ASC + PBS and ASC + CM groups were containing ASCs of 5.0 × 106 cells/ml. We analyzed the treatment of 4 groups to scar tissue after 2 and 4 weeks by hematoxylin and eosin stain, immunohistochemistry, and RNA expression level of tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), and matrix metalloproteinase-2 (MMP-2). Also, the expression of keratin 16 (K16) was detected by western blot analysis. H&E stain showed that infiltration of inflammation cells was significantly reduced at 2 and 4 weeks, as well as re-epithelialization was improved in the ASC + CM group. The ASC + CM gourp was reduced both expression levels of TNF-α, IL-1α, and MMP-2 and K16 protein level. In conclusion, the ASCs with CM has a significant curative effect on acne vulgaris scar, more to the point, the CM has a key role on treatment. It could be applied to a therapeutic approach to regenerate to treat acne vulgaris scar.
Acne Vulgaris ; Adipose Tissue ; Adult ; Blotting, Western ; Cicatrix ; Culture Media, Conditioned ; Ear ; Eosine Yellowish-(YS) ; Hematoxylin ; Humans ; Immunohistochemistry ; Inflammation ; Injections, Subcutaneous ; Keratin-16 ; Male ; Matrix Metalloproteinase 2 ; Methods ; Models, Animal ; Necrosis ; New Zealand ; Rabbits ; Re-Epithelialization ; RNA ; Scapula ; Stem Cells

In keloids, the mechanism underlying the excessive accumulation of extracellular matrix after injury of the skin is unclear, and there is no effective treatment because of the incomplete understanding of their pathogenesis; thus, a high recurrence rate is observed. We studied a new marker of keloids to determine a new treatment strategy. First, the keloid gene expression profile (GSE44270) was analyzed (downloaded from the Gene Expression Omnibus database) and the new keloid marker candidate, epidermal growth factor (EGF)-like repeats and discoidin I-like domains 3 (EDIL3) which were upregulated in keloid samples was identified. Knockdown of EDIL3 is known to suppresses angiogenesis by downregulating relevant inhibitory factors that can limit the supply of survival factors to tumor cells from the circulation via the vascular endothelial cells. In keloids, the mechanism of action of EDIL3 may be similar to that in tumors; the inhibition of apoptosis in tumor cells via a reduction in the apoptosis of blood vessels by upregulating an angiogenic factor. To determine whether EDIL3 is involved in keloid formation, we performed knockdown of EDIL3 in keloid fibroblasts in vitro by transfection with anti-EDIL3 small interfering RNA (via microporation). EDIL3 was upregulated in keloid fibroblasts compared with normal fibroblasts in collagen type I, II and III. Our results indicate the control of EDIL3 expression may be a new promising treatment of keloid disease also the molecular targeting of EDIL3 may improve the quality of treatment and reduce the formation of keloids.
Angiogenesis Inducing Agents ; Apoptosis ; Blood Vessels ; Cicatrix* ; Collagen ; Collagen Type I ; Endothelial Cells ; Epidermal Growth Factor* ; Extracellular Matrix ; Fibroblasts ; Gene Expression ; In Vitro Techniques ; Keloid* ; Recurrence ; RNA, Small Interfering ; Skin ; Transcriptome ; Transfection

Inflammatory bowel disease is associated with extraintestinal manifestations involving almost every organ system in the body. Crohn's disease (CD) appears to be more commonly associated with an inflammatory myopathy than ulcerative colitis. However, myopathy of the thigh in patients with CD is rare. We report an unusual site of necrotizing myositis in a patient with CD. A 23-year-old woman presented with swelling and pain at the left popliteal area that had lasted for 1 week. Twenty-two months before admission, she had presented with pyoderma gangrenosum on the left upper chest and was diagnosed with CD. A magnetic resonance imaging scan of her leg revealed diffuse swelling in the left semimembranous muscle and biceps femoris muscle that was compatible with myositis, and a cystic lesion in the distal portion of the semimembranous muscle. The findings from semimembranous muscle biopsy were also consistent with necrotizing myositis. In conclusion, myositis, although rare, can be an extraintestinal manifestation of CD.
Biopsy ; Colitis, Ulcerative ; Crohn Disease* ; Female ; Humans ; Inflammatory Bowel Diseases ; Leg ; Magnetic Resonance Imaging ; Muscles ; Muscular Diseases ; Myositis* ; Pyoderma Gangrenosum ; Thigh ; Thorax ; Young Adult

A capture enzyme-linked immunosorbent assay (capture ELISA) was developed to detect Clostridium botulinum neurotoxin type A (BoNT/A) in assay buffer and human serum. The assay is based upon affinity-purified rabbit polyclonal and biotinylated monoclonal antibodies directed against the BoNT/A complex purified from C. botulinum ATCC19397. For the capture ELISA, the optimized amount (2 microgram/ml) of rabbit polyclonal antibody was immobilized on ELISA plates to detect BoNT/A (ranging from 0 to 500 ng/ml), which was recognized by 2 microgram/ml of the monoclonal antibody. From three independent repeated experiments, standard curves were linear over the range of 0~31.25 ng/ml BoNT/A and the coefficients (r(2)) ranged from 0.9951~0.9999 for all assays. The inter-variations were typically 0.50~6.93% and the specificity was confirmed by showing no cross-reactivity against BoNT/B and /E. The detection limit of capture ELISA was 0.488 ng/ml, which was close to mouse LD(50). In addition, application with BoNT/A-spiking human sera showed a possibility to detect BoNT/A with capture ELISA from the contaminated human sera. Taken together, the newly developed capture ELISA could serve as a rapid and sensitive screening tool for detecting BoNT/A simultaneously from massive specimens.
Animals ; Antibodies, Monoclonal ; Clostridium botulinum ; Enzyme-Linked Immunosorbent Assay ; Humans ; Limit of Detection ; Mass Screening ; Mice ; Sensitivity and Specificity

BACKGROUND/AIMS: Proton pump inhibitor (PPI)-based standard triple therapy for Helicobacter pylori infection is widely used, but it has a considerable failure rate. The aim of this study was to evaluate the efficacy and tolerability of rescue therapies with a quadruple regimen and a rifabutin-based regimen for patients who experienced failure with PPI-based standard triple therapy. METHODS: From July 2004 through October 2006, 52 patients for whom first line triple therapy (PPI, amoxicillin and clarithromycin) had failed were included in this study. They were treated with a quadruple regimen for 7 days (PPI, bismuth, tetracycline and metronidazole) as a second line therapy. For third line therapy, a rifabutin-based regimen (PPI, rifabutin and amoxicillin) was prescribed for 14 days. The H. pylori status was determined before and at least 4 weeks after therapy by the 13C urea breath test or by endoscopy with antral and corpus biopsies for a rapid urease test, histological examination and culture. RESULTS: The mean age was 52.6 years. Thirteen patients (25%) of the 52 patients were dropped. The eradication rate of the quadruple therapy was 84.6% (33/39). Three patients of the 6 failures with quadruple therapy were then treated with the rifabutin-based regimen. The eradication rate of the rifabutin-based therapy was 100% (3/3). Adverse effects (10.2%) were reported in 4 patients who were treated with quadruple regimen. CONCLUSIONS: The quadruple regimen is still an effective second-line therapy for Korean patients who experience failure with PPI-based standard triple therapy. The rifabutin-based regimen could be used as a third-line rescue therapy in Korea.
Amoxicillin ; Biopsy ; Bismuth ; Breath Tests ; Carbamates ; Endoscopy ; Helicobacter ; Helicobacter pylori ; Humans ; Organometallic Compounds ; Proton Pumps ; Protons ; Rifabutin ; Tetracycline ; Urea ; Urease

No abstract availble.
Anti-Bacterial Agents/therapeutic use ; Disk Diffusion Antimicrobial Tests ; *Drug Resistance, Bacterial ; Drug Therapy, Combination ; Helicobacter Infections/*drug therapy ; *Helicobacter pylori/drug effects/isolation & purification ; Humans ; Microbial Sensitivity Tests ; Peptic Ulcer/*drug therapy

Cytomegalovirus (CMV) colitis usually affects immunosuppressed patients. However, CMV colitis may also affect patients with a debilitation caused by a severe illness or affect patients that have a specific physiological status (old age, pregnancy). Clinically, patients with CMV colitis most commonly present with abdominal pain, diarrhea, and gastrointestinal bleeding. The diagnosis of CMV colitis usually requires a biopsy of mucosal tissue. The characteristic finding on biopsy reveals CMV inclusion bodies. CMV colitis can be successfully treated with ganciclovir. We report two cases of CMV proctocolitis in elderly patients with chronic diseases that presented with massive lower gastrointestinal bleeding due to multiple rectal ulcerations. A pathological examination showed CMV inclusion bodies. CMV colitis should be considered in the differential diagnosis of patients with massive rectal ulcer bleeding when other causes fail to explain the course of the disease.
Abdominal Pain ; Aged ; Biopsy ; Chronic Disease ; Colitis ; Cytomegalovirus* ; Diagnosis ; Diagnosis, Differential ; Diarrhea ; Ganciclovir ; Hemorrhage* ; Humans ; Inclusion Bodies ; Mucous Membrane ; Proctocolitis* ; Ulcer