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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.
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BACKGROUND@#Angiogenesis and vasculogenesis are essential processes for successful tissue regeneration in tissue engineering and regenerative medicine. The adipose-derived stromal vascular fraction (SVF) is not only a source of adipose stem cells (ASC) but also a suitable source of microvascular endothelial cells because it is a rich capillary network. So, we propose a new hypothesis for isolating adipose-derived human microvascular endothelial cells (HMVEC-A) from the SVF and developed a dual isolation system that isolates two cell types from one tissue.METHOD: To isolate HMVEC-A, we analyzed the supernatant discarded when ASC is isolated from the adipose-derived SVF. Based on this analysis, we assumed that the SVF adherent to the bottom of the culture plate was divided into two fractions: the stromal fraction as the ASC-rich fraction, and the vascular fraction (VF) as the endothelial cells-rich fraction floating in the culture supernatant. VF isolation was optimized and the efficiency was compared, and the endothelial cells characteristics of HMVEC-A were confirmed by flow cytometric analysis, immunocytochemistry (ICC), a DiI-acetylated low-density lipoprotein (DiI-Ac-LDL) uptake, and in vitro tube formation assay. @*RESULTS@#Consistent with the hypothesis, we found a large population of HMVEC-A in the VF and isolated these HMVEC-A by our isolation method. Additionally, this method had higher yields and shorter doubling times than other endothelial cells isolation methods and showed typical morphological and phenotypic characteristics of endothelial cells. @*CONCLUSION@#Cells obtained by the method according to our hypothesis can be applied as a useful source for studies such as tissue-to-tissue networks, angiogenesis and tissue regeneration, patient-specific cell therapy, and organoid chips.
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BACKGROUND@#Verapamil is used in the treatment of hypertension, angina pectoris, cardiac arrhythmia, hypertrophic scars, and keloids to block transmembrane calcium ion flux. Verapamil has antioxidant activity, which enhances the production of nitric oxide (NO). NO promotes the proliferation of fibroblasts, keratinocytes, endothelial cells, and epithelial cells during wound healing. In this study, we investigated the effect of verapamil and its antioxidant properties on the enhancement of acute wound healing via NO. @*METHODS@#A full-thickness wound healing model was created on the rat dorsal with a silicone ring. The wound closure rate was estimated every 2 days for 14 days. A histological study was performed to evaluate wound healing.Immunofluorescence staining was analyzed for angiogenesis. The expressions of collagen type I (COL I), collagen type III (COL III), and vascular endothelial growth factor (VEGF) were assessed by Western blot. Real-time polymerase chain reaction (qRT-PCR) was performed to examine the expression of endothelial NO synthase and inducible NO synthase, which are related to antioxidant activity in the process of wound healing. @*RESULTS@#The wound closure rate was faster in the verapamil group compared to the control and silicone groups.Histologic analysis revealed capillaries and stratum basale in the verapamil group. Immunofluorescence staining was shown vessel formation in the verapamil group. Western blot and qRT-PCR analysis revealed high expression levels of COL I, VEGF, eNOS, and FGF in the verapamil. @*CONCLUSION@#Verapamil’s antioxidant activity enhances NO production in acute wound healing. We suggest that verapamil can be used to promote acute wound healing.
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BACKGROUND@#Angiogenesis and vasculogenesis are essential processes for successful tissue regeneration in tissue engineering and regenerative medicine. The adipose-derived stromal vascular fraction (SVF) is not only a source of adipose stem cells (ASC) but also a suitable source of microvascular endothelial cells because it is a rich capillary network. So, we propose a new hypothesis for isolating adipose-derived human microvascular endothelial cells (HMVEC-A) from the SVF and developed a dual isolation system that isolates two cell types from one tissue.METHOD: To isolate HMVEC-A, we analyzed the supernatant discarded when ASC is isolated from the adipose-derived SVF. Based on this analysis, we assumed that the SVF adherent to the bottom of the culture plate was divided into two fractions: the stromal fraction as the ASC-rich fraction, and the vascular fraction (VF) as the endothelial cells-rich fraction floating in the culture supernatant. VF isolation was optimized and the efficiency was compared, and the endothelial cells characteristics of HMVEC-A were confirmed by flow cytometric analysis, immunocytochemistry (ICC), a DiI-acetylated low-density lipoprotein (DiI-Ac-LDL) uptake, and in vitro tube formation assay. @*RESULTS@#Consistent with the hypothesis, we found a large population of HMVEC-A in the VF and isolated these HMVEC-A by our isolation method. Additionally, this method had higher yields and shorter doubling times than other endothelial cells isolation methods and showed typical morphological and phenotypic characteristics of endothelial cells. @*CONCLUSION@#Cells obtained by the method according to our hypothesis can be applied as a useful source for studies such as tissue-to-tissue networks, angiogenesis and tissue regeneration, patient-specific cell therapy, and organoid chips.
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BACKGROUND@#Verapamil is used in the treatment of hypertension, angina pectoris, cardiac arrhythmia, hypertrophic scars, and keloids to block transmembrane calcium ion flux. Verapamil has antioxidant activity, which enhances the production of nitric oxide (NO). NO promotes the proliferation of fibroblasts, keratinocytes, endothelial cells, and epithelial cells during wound healing. In this study, we investigated the effect of verapamil and its antioxidant properties on the enhancement of acute wound healing via NO. @*METHODS@#A full-thickness wound healing model was created on the rat dorsal with a silicone ring. The wound closure rate was estimated every 2 days for 14 days. A histological study was performed to evaluate wound healing.Immunofluorescence staining was analyzed for angiogenesis. The expressions of collagen type I (COL I), collagen type III (COL III), and vascular endothelial growth factor (VEGF) were assessed by Western blot. Real-time polymerase chain reaction (qRT-PCR) was performed to examine the expression of endothelial NO synthase and inducible NO synthase, which are related to antioxidant activity in the process of wound healing. @*RESULTS@#The wound closure rate was faster in the verapamil group compared to the control and silicone groups.Histologic analysis revealed capillaries and stratum basale in the verapamil group. Immunofluorescence staining was shown vessel formation in the verapamil group. Western blot and qRT-PCR analysis revealed high expression levels of COL I, VEGF, eNOS, and FGF in the verapamil. @*CONCLUSION@#Verapamil’s antioxidant activity enhances NO production in acute wound healing. We suggest that verapamil can be used to promote acute wound healing.
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Background@#Nipple reconstruction is usually performed as a delayed procedure in patients with breast cancer who undergo skin-sparing mastectomy and breast reconstruction surgery using a deep inferior epigastric perforator (DIEP) flap. The authors designed this study to evaluate the utility of breast reconstruction based on a DIEP flap and immediate nipple reconstruction. @*Methods@#A retrospective review was conducted of all patients who underwent breast reconstruction performed by a single plastic surgeon from October 2016 to June 2018. Through a questionnaire and chart review, we compared surgical results and complications in cases of single-stage nipple reconstruction after skin-sparing mastectomy (n=17) with patients who underwent delayed nipple reconstruction after skin-sparing mastectomy, modified radical mastectomy, or simple mastectomy (n=7). @*Results@#In a subjective analysis using clinical photos, the immediate nipple reconstruction group had higher scores than their counterparts in an evaluation of the nipple-areolar complex (NAC) (NAC placement, 3.34 vs. 3.04; nipple projection, 3.05 vs. 3.03; nipple size, 3.30 vs. 3.29). No significant differences between the groups were found in terms of complications. @*Conclusions@#Simultaneous nipple reconstruction is a reliable surgical method with economic advantages. No differences were found in terms of outcomes and complications in comparison to delayed reconstruction. Therefore, surgeons can consider simultaneous nipple reconstruction without particular concerns about asymmetry or necrosis.
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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.
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BACKGROUND: Silica particles (SPs) induce cell proliferation and osteogenic differentiation. We reported that SPs in the scaffold induced early stage osteogenic differentiation. METHODS: A polycaprolactone (PCL) scaffold was fabricated with a 10 wt% SPs. The surface of PCL scaffold was coated with a 10 µg/mL collagen solution. Next, the scaffold was conjugated with 2 µM SPs, 2 µg/mL bone morphogenetic protein 2 (BMP2), or 2 µM BMP2-conjugated SPs (BCSPs). Green fluorescent protein-coupled BMP2 was applied to fabricate the scaffold. The fluorescence intensity was analyzed by confocal microscopy. The mRNA levels of the early osteogenic differentiation marker, alkaline phosphatase (ALP), were analyzed by real-time quantitative polymerase chain reaction. Levels of BMP2, RUNX2, ERK1/2, and AKT were assessed by western blotting. RESULTS: ALP mRNA levels were significantly higher in the BCSP-conjugated scaffold than in the other scaffolds. In the early stage of osteogenic differentiation, the protein levels of BMP2, RUNX2, ERK1/2, and AKT in cells were significantly higher in the BCSP-conjugated scaffold than in other scaffolds. Thus, the BCSP composite scaffold induced rapid osteogenic differentiation. CONCLUSION: These results suggest that BCSP composite can be used to promote early stage osteogenic differentiation and show promise as a material for use in scaffolds for bone regeneration.
Subject(s)
Alkaline Phosphatase , Blotting, Western , Bone Morphogenetic Protein 2 , Bone Morphogenetic Proteins , Bone Regeneration , Cell Proliferation , Collagen , Fluorescence , Microscopy, Confocal , Polymerase Chain Reaction , RNA, Messenger , Silicon Dioxide , Stem CellsABSTRACT
Salivary gland tumors usually appear in solitary mass in single salivary gland. The coexistence of tumors with different histological types occurring within a unilateral parotid gland is an extremely rare event. We experienced a case which two different types of malignant tumors developed at different time points in same gland; metachronous tumors. The second tumor was excised widely and reconstruction was performed by free tissue transfer. Sensory and motor nerve to the left cheek appeared to be intact, and circulation was adequate. This rare case was presented in this article.
Subject(s)
Adenocarcinoma , Cheek , Neoplasms, Second Primary , Parotid Gland , Salivary GlandsABSTRACT
Macroglossia is a rare clinical condition defined as an enlarged tongue. Macroglossia can cause structural deformities like diastema and disproportionate mandibular growth and present functional disorders such as dysarthria, dysphonia, and respiratory problems. A 7-year-old boy who had lymphangiomatous macroglossia was treated with a reduction glossectomy by anchor-shaped combination of a U-shape and modified key-hole resection. Postoperatively, the reduced tongue was contained completely within the oral cavity, but open bite remained due to prognathism. Sensory and motor nerves to the tongue appeared to be intact, and circulation was adequate. This patient will be monitored for recurrence of tongue enlargement.
Subject(s)
Child , Humans , Male , Congenital Abnormalities , Diastema , Dysarthria , Dysphonia , Glossectomy , Lymphangioma , Macroglossia , Mouth , Open Bite , Prognathism , Recurrence , TongueABSTRACT
BACKGROUND: Various methods have been reported for the improvement of widened or depressed scars. However, scars can be complex at presentation. If a widened but flat portion is combined with a tethered or a depressed area, a scar revision method that can effectively resolve all issues is needed. The authors utilized a dermal portion of the widened scar as a re-adhesion barrier and filled tissue after the release of the tethering or depression. METHODS: From July 2014 to December 2017, a total of eight patients presented with combined scars with both widened and depressed areas and underwent scar revisions with scar dermal transposition flaps. The scar flap of the widened scar was de-epithelialized, leaving the dermo-cutaneous pedicle near the depressed area. Without any additional skin incision on the depressed site, the tethering was released by making a skin incision at the scar flap. The de-epithelialized scar dermal flap was transpositioned under the depressed area of the scar. RESULTS: The surgical wounds of all eight patients healed without any complications. The mean follow-up period was 5.25 months. The filling effect of the scar flap persisted without the conspicuous relapse of a depression or tethering. The patients were satisfied with the final results and the fact that no additional incision was needed for the tethered and depressed scar. CONCLUSIONS: If the depressed site is near a widened scar, a scar dermal transposition flap may be a versatile option for improving the depression without an additional skin incision.
Subject(s)
Humans , Autografts , Cicatrix , Depression , Follow-Up Studies , Methods , Recurrence , Skin , Surgical Flaps , Wounds and InjuriesABSTRACT
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.
Subject(s)
Adult , Humans , Male , Rabbits , Acne Vulgaris , Adipose Tissue , Blotting, Western , Cicatrix , Culture Media, Conditioned , Ear , Eosine Yellowish-(YS) , Hematoxylin , Immunohistochemistry , Inflammation , Injections, Subcutaneous , Keratin-16 , Matrix Metalloproteinase 2 , Methods , Models, Animal , Necrosis , New Zealand , Re-Epithelialization , RNA , Scapula , Stem CellsABSTRACT
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.
Subject(s)
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 , TransfectionABSTRACT
BACKGROUND: Creating a natural-looking umbilicus during closure of the donor-site in abdominally based free flap breast reconstruction is a factor of satisfaction for both the patient and surgeon. We present a simple method of umbilical transposition that results in an aesthetic, natural-looking umbilicus. METHODS: From March 2011 to November 2014, fifty three consecutive female patients received abdominal flap breast reconstruction. Twenty patients (from March 2011 to February 2013) underwent umbilical transposition through a cross like incision in the abdominal flap, with fascial fixation sutures but no dermal flaps. Thirty three patients (March 2013 to November 2014) received umbilical transposition in the following method. An oval-shaped incision is made at the location of the new umbilicus on the abdominal flap. This oval is deepithelialized, and full-thickness incisions are made at the 2, 6, and 10 o'clock directions to create three triangular dermal flaps. These are pulled down to the abdominal fascia using sutures that pass through the umbilical stalk and the abdominal fascia at the 3, 9, and 12 o'clock directions. This results in an umbilical stalk lined with dermal flaps, creates a natural periumbilical concavity, and anchors the abdominal flap inward to minimize tension. The cranial flap enhances superior hooding. RESULTS: Patient and surgeon satisfaction, surveyed 2 months after surgery with a satisfaction scoring system, were higher in the dermal flap group. CONCLUSIONS: The technique using three dermal flaps in an oval skin incision is simple, relatively easy to learn, and results in an aesthetic, natural-looking umbilicus.
Subject(s)
Female , Humans , Abdominoplasty , Fascia , Free Tissue Flaps , Mammaplasty , Methods , Skin , Surgical Flaps , Sutures , UmbilicusABSTRACT
Mesenchymal stromal cells (MSCs) established by in-vitro adherence culture have been widely utilized for various cell therapeutic trials, but potential heterogeneity that can be caused by preparation methods are poorly characterized. In this study, we show that at least two distinct subsets of MSCs with different adherence to plastic surface exist in human adipose tissue-derived stromal vascular fraction (SVF); while 69% of total colony forming units in SVF adhere to the surface before 3 hrs of plating, 13–17% of colonogenic cells adhered to the surface at later period of 15 hr to 1 week after plating. Of note, the late adherent MSCs exhibited higher self-renewal of colony forming cells and higher proliferating potential with comparable level of osteogenic or adipogenic differentiation potential to the early adherence subsets. Moreover, late adherent cells exhibited distinct pattern of paracrine secretome including higher level secretion of cytokines than the early adherent subsets. Taken together, these results suggest the possibility that distinct adherence properties of MSCs can be another parameter of clonal heterogeneity in the subpopulations of adipose tissue MSCs and that it can be an important factor for optimization of MSC preparation for cell therapeutic trials.
Subject(s)
Humans , Adipose Tissue , Cytokines , Mesenchymal Stem Cells , Plastics , Population Characteristics , Stem CellsABSTRACT
BACKGROUND: The prenatal ultrasound detection of cleft lip with or without cleft palate (CL/P) and its continuous management in the prenatal, perinatal, and postnatal periods using a multidisciplinary team approach can be beneficial for parents and their infants. In this report, we share our experiences with the prenatal detection of CL/P and the multidisciplinary management of this malformation in our institution's Congenital Disease Center. METHODS: The multidisciplinary team of the Congenital Disease Center for mothers of children with CL/P is composed of obstetricians, plastic and reconstructive surgeons, pediatricians, and psychiatrists. A total of 11 fetuses were diagnosed with CL/P from March 2009 to December 2013, and their mothers were referred to the Congenital Disease Center of our hospital. When CL/P is suspected in the prenatal ultrasound screening examination, the pregnant woman is referred to our center for further evaluation. RESULTS: The abortion rate was 28% (3/11). The concordance rate of the sonographic and final diagnoses was 100%. Ten women (91%) reported that they were satisfied with the multidisciplinary management in our center. CONCLUSIONS: Although a child with a birth defect is unlikely to be received well, the women whose fetuses were diagnosed with CL/P on prenatal ultrasound screening and who underwent multidisciplinary team management were more likely to decide to continue their pregnancy.
Subject(s)
Child , Female , Humans , Infant , Pregnancy , Abortion, Induced , Cleft Lip , Cleft Palate , Congenital Abnormalities , Craniofacial Abnormalities , Diagnosis , Fetus , Interdisciplinary Communication , Mass Screening , Mothers , Palate , Parents , Plastics , Pregnant Women , Prenatal Care , Psychiatry , UltrasonographyABSTRACT
BACKGROUND: The erbium:yttrium scandium gallium garnet (Er:YSGG) laser differs from other laser techniques by having a faster and higher cure rate. Since the Er:YSGG laser causes an appropriate proportion of ablation and coagulation, it has advantages over the conventional carbon dioxide (CO2) laser and the erbium-doped yttrium aluminum garnet (Er:YAG) laser, including heating tendencies and explosive vaporization. This research was conducted to explore the effects and safety of the Er:YSGG laser. METHODS: Twenty patients participated in the pilot study of a resurfacing system using a 2,790-nm Er:YSGG laser. All patients received facial treatment by the 2,790-nm Er:YSGG laser system (Cutera) twice with a 4-week interval. Wrinkle reduction, reduction in pigment inhomogeneity, and improvement in tone and texture were measured. RESULTS: Study subjects included 15 women and five men. Re-epithelization occurred in all subjects 3 to 4 days after treatment, and wrinkle reduction, reduction in pigment inhomogeneity, and improvement in tone and texture within 6 months of treatment. CONCLUSIONS: The 2,790-nm YSGG laser technique had fewer complications and was effective in the improvement of scars, pores, wrinkles, and skin tone and color with one or two treatments. We expect this method to be effective for people with acne scars, pore scars, deep wrinkles, and uneven skin texture and color.
Subject(s)
Female , Humans , Male , Ablation Techniques , Acne Vulgaris , Aluminum , Carbon Dioxide , Cicatrix , Gallium , Heating , Hot Temperature , Laser Therapy , Lasers, Solid-State , Pilot Projects , Rejuvenation , Scandium , Skin , Volatilization , YttriumABSTRACT
No abstract available.