RÉSUMÉ
Purpose: Anal wounds following hemorrhoidectomy can lead to severe pain and postoperative bleeding, impacting patient recovery and quality of life. Hyaluronic acid (HA) stimulates tissue regeneration and wound healing by accelerating cell migration and proliferation. This study aimed to investigate the differences in wound healing rate and completeness of recovery of perianal wounds topically treated with HA-soaked cotton in a murine model. Methods: Forty-eight 8-week-old Sprague-Dawley rats with perianal wounds created using a biopsy punch were divided into 2 groups: simple dressing with gauze (control) and topical HA-soaked cotton. A single application of HA-soaked cotton was administered after surgery. Wound healing rate and completeness of recovery were evaluated by measuring the healed area and conducting histological analyses. Results: The HA-cotton group exhibited a shorter complete wound healing duration compared to the control group (13.9 days vs. 16.4 days, P = 0.031). Differences in wound healing area between the 2 groups were greatest on postoperative day 2 (51.6% vs. 28.8%, P < 0.001). The HA-cotton group exhibited fewer cases of granulation tissue (2 vs. 5) or redness (0 vs. 3) upon complete wound healing. Histologically, the HA-cotton group showed accelerated reepithelialization, rapid shift to lymphocyte-dominant inflammation, enhanced fibroblast proliferation, and increased collagen deposition compared to the control group. Conclusion: Herein, topical application of HA-soaked cotton on perianal wounds in rats resulted in accelerated wound healing, particularly in the initial stages, and improved completeness of recovery, underscoring the potential of the topical application of HA-soaked cotton on hemorrhoidectomy wounds in human patients to improve wound healing.
RÉSUMÉ
INTRODUCTION: Although various products are commonly used for skin rejuvenation, solid-type hyaluronic acid (HA) as an injectable form has not been researched or utilized. This study aimed to demonstrate the safety and efficacy of solid-type HA in thread form, which differs from the conventional gel-type HA commonly used. METHOD: Solid-type HA threads, conventional HA fillers, and polydioxanone (PDO) threads were inserted into the dorsal subcutaneous layer of mice. Photographs were taken on days 0, 1, 3, and 7, and on day 7, the samples were harvested for histological analysis. Inflammatory reactions and detection of collagen were confirmed through tissue staining, and real-time PCR was conducted to quantify collagen synthesis. RESULTS: In the histological analysis, the PDO threads exhibited a greater inflammatory response compared to the HA threads. Masson's trichrome staining revealed a higher degree of collagen synthesis in the HA thread group compared to the HA filler group. While collagen type 1 expression was significantly higher in the PDO thread group than in the HA thread group, the HA thread group showed higher expression levels of collagen type 3. Furthermore, the PDO thread group demonstrated a statistically significant increase in TGF-ß1 compared to the HA group. CONCLUSION: This in vivo study demonstrated the stable application of solid-type pure HA threads and their potential for inducing collagen production, while also yielding a low inflammatory response. The findings highlight the promising applications of solid-type HA in the field of cosmetic dermatology. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .
Sujet(s)
Produits de comblement dermique , Souris , Animaux , Produits de comblement dermique/effets indésirables , Polydioxanone , Acide hyaluronique/effets indésirables , Peau , CollagèneRÉSUMÉ
The oral route is considered an attractive method for drug delivery, as it avoids the hepatic and intestinal first-pass metabolism processes. Hyaluronic acid (HA) beneficial effects to the human body include anti-aging and wound healing but its effects on oral barrier integrity and mechanical function have not yet been investigated. In this study, we analyzed oral barrier integrity and the paracellular pathway of HA transportation in TR146 cell monolayers during and after permeation and using low molecular weight HA (LMW-HA, <100 kDa) and high molecular weight HA (HMW-HA, >500 kDa). Cytotoxicity assays in TR146 cells revealed that neither LMW-HA or HMW-HA altered cell viability at concentrations <0.5 % during 24 h of treatment. HA-treated TR146 cell monolayers showed enhanced oral barrier integrity and reduced apparent permeability of fluorescein. Moreover, HA significantly increased tight junction (TJ)-related genes expression, including ZO-2, marvelD3, cingulin, claudin-1, claudin-3, and claudin-4 expression. Overall, the results of the present study indicate that HA can permeate across the oral barrier and enhance oral barrier function via the upregulated expression of TJ-related genes.
Sujet(s)
Acide hyaluronique , Jonctions serrées , Humains , Acide hyaluronique/pharmacologie , Acide hyaluronique/métabolisme , Masse moléculaire , Perméabilité , Muqueuse intestinale/métabolisme , Cellules Caco-2RÉSUMÉ
Background/purpose: Oral wound healing undergoes a dynamic process of oral environment. This study aimed to investigate the effects of hyaluronic acid (HA) film on oral wound healing in a rat model. Materials and methods: A total of 60 rats with tongue wounds (5 mm in diameter) were randomly divided into control (n = 20), HA gel (n = 20), and HA film groups (n = 20). The animals were sacrificed on either 3 or 7 days after the experiment. Clinical, histological, and quantitative reverse transcriptase-polymerase chain reaction analysis were performed to evaluate the healing rate, inflammation, re-epithelialization, and gene expression of wound healing biomarkers. Results: The healing rates of HA gel (84.4 ± 9.2%) and HA film (74.0 ± 15.0%) were significantly higher than that of the control (51.7 ± 16.9%) on day 7 (P < 0.001 and P = 0.002, respectively). Histological analysis revealed no significant differences between the groups on day 3. On day 7, only the HA film showed significant improvement in inflammation (P = 0.038) and re-epithelialization (P = 0.011) compared to the control. Regarding wound healing biomarkers, both HA gel and HA film groups showed lower level of COL1α1 expression on day 3 compared to the control. Conclusion: Within the limits of this study, HA film was found to be effective for oral wound healing, particularly for re-epithelialization. This finding suggests that HA film delivery can be beneficial not only for clinical convenience but also for promoting oral wound healing.
RÉSUMÉ
PURPOSE: Postoperative pain and delayed wound healing are the main complications following anal surgery associated with poor quality of life. Hyaluronic acid (HA) supports tissue regeneration and rapid wound healing by promoting cell proliferation and migration. We investigated the effects of HA on perianal wound healing in a rat model. METHODS: Forty-eight 8-week-old Sprague-Dawley rats with perianal wounds created by biopsy punch were divided into 3 groups: simple dressing with gauze (control), dressing with topical HA film, and dressing with topical HA gel. HA agents were not reapplied postoperatively. Wound healing was evaluated by measuring the healed area, and histological analyses were randomly performed using hematoxylin and eosin and Masson trichrome staining. RESULTS: Fewer mean days were required for complete wound healing in the HA film and HA gel groups than in the control group (11.6 vs. 11.9 vs. 13.8 days, respectively; P = 0.010). The healed area in the HA film group on day 11 was larger than that in the HA gel and control groups (80.2% vs. 61.9% vs. 53.2%, respectively; P < 0.001). Histologically, the HA film group showed accelerated reepithelialization, a rapid transition to lymphocyte-predominant inflammation, and increased fibroblastic proliferation and collagen deposition compared to the other groups. There was no treatment-related toxicity in the HA application groups. CONCLUSION: Topical application of HA film to perianal wounds improves the wound healing rate in a rat model. This finding suggests a potential benefit of HA film application in promoting wound healing after anal surgery in humans.
RÉSUMÉ
Orodispersible films (ODF) were prepared with mixtures of hyaluronic acid (HA) and carboxymethyl cellulose (CMC), and the effect of CMC addition on the disintegration and mechanical properties of the composite films were examined. Low molecular weight HA (10 kDa) appeared more acceptable for ODF than high molecular weight HA (800 kDa) because of its rapid disintegration in the oral cavity. The composite films appeared similar to pullulan film with excellent transparency and surface smoothness. The disintegration time as well as mechanical properties of the films such as tensile strength and elongation at break were increased by the addition of CMC. Overall, the CMC addition, up to 35%, improved the mechanical properties of low molecular weight HA film within a proper range of disintegration time for ODF.
RÉSUMÉ
The regulation of mitochondrial biogenesis, melanogenesis, and connective tissue proteins is critical for homeostasis and aging skin cells. We examined the biological effects of polydeoxyribonucleotide (PDRN) on mitochondrial biogenesis, melanogenesis, and connective tissue proteins in vitro. In a radical scavenging assay, PDRN showed antioxidant activities in a dose-dependent manner, and those activities can suppress cellular oxidative stress in skin cells. PDRN directly inhibited mushroom tyrosinase activity and cellular tyrosinase activity, thus significantly reducing the cellular melanin content in B16-F10 melanocytes. The mRNA and protein expressions of the microphthalmia-associated transcription factor (MITF), which is a key melanogenic gene transcription factor, were significantly downregulated by PDRN. Accordingly, tyrosinase-related protein 1, dopachrome tautomerase, and tyrosinase, which gene expressions were regulated by MITF, were significantly downregulated by PDRN. Mitotracker-probed mitochondria image analysis suggested that PDRN enhanced mitochondrial density in both murine melanoma cells and in human skin fibroblast cells. In addition, PDRN strongly suppressed in vitro elastase enzyme activity in a dose-dependent manner and inhibited matrix metalloproteinase-1 gene expression in human skin fibroblast cells. Collectively, these findings indicate that PDRN has multiple beneficial biological activities in skin cells: hypopigmentation, induction of mitochondrial biogenesis, and the inhibition of collective tissue proteins.
Sujet(s)
Matrix metalloproteinase 1/métabolisme , Mélanines/biosynthèse , Mitochondries/effets des médicaments et des substances chimiques , Biogenèse des organelles , Polydésoxyribonucléotides/pharmacologie , Peau/métabolisme , Animaux , Lignée cellulaire , Régulation négative , Fibroblastes , Régulation de l'expression des gènes/effets des médicaments et des substances chimiques , Humains , Mélanines/génétique , Souris , Facteur de transcription associé à la microphtalmie/métabolisme , Oxidoreductases/métabolisme , ARN messager/métabolismeRÉSUMÉ
This study presents laboratory methods for the quantification of hypopigmentation activity in vitro. Melanin, the major pigment in melanocytes, is synthesized in response to multiple cellular and environmental factors. Melanin protects skin cells from ultraviolet damage, but also has biophysical and biochemical functions. Excessive production or accumulation of melanin in melanocytes can cause dermatological problems, such as freckles, dark spots, melasma, and moles. Therefore, the control of melanogenesis with hypopigmentation agents is important in individuals with clinical or cosmetic needs. Melanin is primarily synthesized in the melanosomes of melanocytes in a complex biochemical process called melanogenesis, which is influenced by extrinsic and intrinsic factors, such as hormones, inflammation, age, and ultraviolet light exposure. We describe three methods to determine the hypopigmentation activity of chemicals or natural substances in melanocytes: measurement of the 1) cellular tyrosinase activity and 2) melanin content, and 3) staining and quantifying cellular melanin with image analysis. In melanogenesis, tyrosinase catalyzes the rate-limiting step that converts L-tyrosine into 3,4-dihydroxyphenylalanine (L-DOPA) and then into dopaquinone. Therefore, the inhibition of tyrosinase is a primary hypopigmentation mechanism. In cultured melanocytes, tyrosinase activity can be quantified by adding L-DOPA as a substrate and measuring dopaquinone production by spectrophotometry. Melanogenesis can also be measured by quantifying the melanin content. The melanin-containing cellular fraction is extracted with NaOH and melanin is quantified spectrophotometrically. Finally, the melanin content can be quantified by image analysis following Fontana-Masson staining of melanin. Although the results of these in vitro assays may not always be reproduced in human skin, these methods are widely used in melanogenesis research, especially as the initial step to identify potential hypopigmentation activity. These methods can also be used to assess melanocyte activity, growth, and differentiation. Consistent results with the three different methods ensure the validity of the effects.
Sujet(s)
Hypopigmentation/diagnostic , Animaux , Arbutoside/pharmacologie , Humains , Mélanines/biosynthèse , Mélanocytes/effets des médicaments et des substances chimiques , Mélanocytes/métabolisme , Mélanocytes/anatomopathologie , Tyrosine/métabolismeRÉSUMÉ
Waxy maize starch was dispersed (14.7% solids) in an aqueous sulfuric acid solution (3.16M), and hydrolyzed by stirring for up to 7 days at 40°C with ultrasonic treatments at different vibration amplitudes (20 and 40%) and durations (30 and 60min/day). The amount of starch nanoparticles in the hydrolyzates isolated after 7 days, measured by a dynamic light scattering detector, was raised from 20% to 70% by an ultrasonic treatment (20% amplitude, 30min). The aggregation of nanoparticles possibly occurring during the hydrolysis was effectively prevented by the ultrasonication. Alternatively, ultrasonic treatments were applied to the re-dispersed suspension of the large microparticles of starch hydrolyzates (2 days) precipitated by a mild centrifugation (500rpm, 10min). By an ultrasonic treatment at 60% vibration amplitude for 3min, the microparticles could be completely transformed to nanoparticles. The inherent crystalline structure of waxy maize starch (A-type in X-ray diffraction) remained after the ultrasonic treatments during acid hydrolysis, but it was disrupted by the ultrasonic treatments for the re-dispersed microparticles.
Sujet(s)
Nanoparticules/composition chimique , Nanotechnologie/méthodes , Amidon/composition chimique , Science des ultrasons/méthodes , Zea mays/composition chimique , Acides/composition chimique , Cristallisation , Hydrodynamique , Hydrolyse , Microscopie électronique à balayage , Nanoparticules/ultrastructure , Taille de particule , Amidon/ultrastructure , Suspensions/composition chimique , Température , Facteurs temps , Diffraction des rayons XRÉSUMÉ
Amylosucrase (AS), maltooligosyltrehalose synthase (MTS), and maltooligosyltrehalose trehalohydrolase (MTH) were used in combined cross-linked enzyme aggregates (combi-CLEAs) to achieve one-step bioconversion of sucrose to trehalose. Combi-CLEAs of three enzymes were successfully established with acetone and glutaraldehyde (GA) as a precipitant and a cross-linker, respectively. The optimum enzyme ratio was 8:0.5:0.5 (AS, 4mg:MTS, 0.25mg:MTH, 0.25mg). To improve trehalose production, bovine serum albumin was co-aggregated with enzymes as a proteic feeder. The trehalose production yield of combi-CLEAs was about 8% in each cycle on the basis of substrate added up to 400mM. Finally, the combi-CLEAs used in this experiment showed reusability of about five cycles without any activity loss.
Sujet(s)
Glucosidases/métabolisme , Glucosyltransferases/métabolisme , Saccharose/métabolisme , Tréhalose/métabolisme , Acétone/composition chimique , Animaux , Bovins , Précipitation chimique , Réactifs réticulants/composition chimique , Glucosidases/composition chimique , Glucosyltransferases/composition chimique , Glutaraldéhyde/composition chimique , Sérumalbumine bovineRÉSUMÉ
Disaccharide glycosides synthesised from food grade resources consist of the hydrophilic head group of maltose or lactose and provide better hydrophilic-lipophilic balance (HLB = 12) to the long alkyl chain derived from palm oil (PO) and palm kernel oil (PKO). Maltoside provides more flexibility in the vesicle's membrane because of its low packing density in the bilayer membrane compared to lactoside. The bending of the molecular structure in maltose form a less compact assembly for maltoside, whereas lactose is more linear in shape. Apart from hydrophilic moieties, packing behaviour was also governed by the hydrophobic moieties. PO has higher degree of unsaturation compared to PKO, thus providing higher fluidity in the bilayer membrane. Vesicle with high membrane flexibility is easier to disintegrate and deform to enhance drug penetration into the skin. Results showed that the glycosides delivered vitamin E (VE) into deeper skin layer at least two-fold higher than free VE.
Sujet(s)
Lipides/composition chimique , Absorption cutanée , Animaux , Mâle , Rat WistarRÉSUMÉ
The potential toxicity of fucoidan from Undaria pinnatifida was investigated in vitro and in vivo. By the Ames test, fucoidan showed no mutagenicity up to 500 microL/plate, and inhibited the mutagenicity induced by 4-nitro-quinoline-1-oxide, by up to 71%, compared with controls. In the bone marrow micronucleus test, fucoidan, at all levels tested, did not change the micronucleated polychromatic erythrocyte percentage ratio in mouse bone marrow cells. As an acute in vivo toxicity test, fucoidan from 0 to 2000 mg/kg body weight per day was administered orally to Sprague-Dawley rats for 28 days. No significant toxicological change was induced by fucoidan treatment up to 1000 mg/kg body weight per day in biochemical analyses, hematological analyses, necropsy and liver histopathology. The plasma ALT level was slightly, but significantly, increased in male rats at 2000 mg/kg/day. The consumption of fucoidan from Undaria pinnatifida, up to 1000 mg/kg body weight per day, may be safe in rodents, with no sign of toxicity after up to 28 days of daily administration.
Sujet(s)
Polyosides/toxicité , Undaria/composition chimique , Administration par voie orale , Animaux , Femelle , Mâle , Souris , Souris de lignée ICR , Tests de micronucleus , Taille d'organe , Rats , Rat Sprague-Dawley , Tests de toxicitéRÉSUMÉ
Puerarin is an isoflavone derived from Kudzu roots and has antioxidant and hypocholesterolemic effects; however, its insolubility often limits its biological availability in vivo. Using a novel transglycosylation process, the solubility of puerarin glycosides was increased >100-fold, but it was not known whether these modified puerarin glycosides maintained biological activities. We found that water-soluble puerarin glycosides fully maintained antioxidant activities compared with puerarin assessed by radical scavenging activity, reducing power assay, superoxide dismutase activity, and non-site-specific hydroxyl radical scavenging activity. Both puerarin and its glycosides also significantly reduced low-density lipoprotein (LDL) oxidation. Mice fed with puerarin glycosides (0.1% w/w) showed significantly reduced plasma total cholesterol levels, thus, we further investigated their hypocholesterolemic mechanisms by assessing several key gene expressions both in vitro and in vivo. Puerarin and its glycosides induced multiple changes in hepatic cholesterol metabolism. The LDL receptor promoter activity was increased dose-dependently in puerarin glycosides-treated HepG2 cells. Accordingly, the expression of LDL receptor mRNA and protein were also significantly increased in HepG2 cells and mouse livers. The transcription and translation of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase were down-regulated both in vitro and in vivo. The cholesterol 7alpha-hydroxylase (CYP7A1) mRNA levels were not affected in vitro but significantly up-regulated in the mouse livers. Collectively, our results show that puerarin and its glycosides are biologically fully active isoflavone and have antioxidant and hypocholesterolemic effects in HepG2 cells and in C57BL/6J mice. In the livers, hypocholesterolemic effects of puerarin glycoside may be achieved by multiple mechanisms including increasing LDL uptake, reducing cholesterol biosynthesis, and possibly enhancing cholesterol degradation.
Sujet(s)
Anticholestérolémiants/pharmacologie , Cholestérol/métabolisme , Piégeurs de radicaux libres/pharmacologie , Hétérosides/pharmacologie , Isoflavones/pharmacologie , Foie/effets des médicaments et des substances chimiques , Acyl coenzyme A/génétique , Acyl coenzyme A/métabolisme , Animaux , Anticholestérolémiants/composition chimique , Lignée cellulaire tumorale , Cholestérol/sang , Cholesterol 7-alpha-hydroxylase/génétique , Cholesterol 7-alpha-hydroxylase/métabolisme , Relation dose-effet des médicaments , Piégeurs de radicaux libres/composition chimique , Radicaux libres/composition chimique , Hétérosides/composition chimique , Glycosylation , Hépatocytes/effets des médicaments et des substances chimiques , Hépatocytes/métabolisme , Humains , Isoflavones/composition chimique , Foie/enzymologie , Foie/métabolisme , Souris , Souris de lignée C57BL , ARN messager/métabolisme , Récepteurs aux lipoprotéines LDL/génétique , Récepteurs aux lipoprotéines LDL/métabolisme , Solubilité , Solvants/composition chimique , Transcription génétique/effets des médicaments et des substances chimiques , Transfection , Eau/composition chimiqueRÉSUMÉ
To make effective wound healing accelerator, water-soluble chitosan (WSC)/heparin (CH) complex was prepared using WSC with wound healing ability and heparin with ability to attract or bind growth factor related to wound healing process. Water-soluble CH complex was prepared by the reaction between WSC and heparin, and then, by adding distilled water to it, ointment type with high viscosity was made. To evaluate the wound healing effect, full thickness skin excision was performed on the backs of the rat and then WSC and water-soluble CH complex ointments were applied in the wounds, respectively. After 15 days, gross and histologic examination was performed. Grossly, untreated control group revealed that the wound had well defined margin and was covered by crust. The second group treated with WSC ointment revealed small wound size with less amount of covering crust and ill-defined margin, which appeared to regenerate from margin. The third group treated with water-soluble CH complex ointment appeared to be nearly completely healed. Histology of each group was well correlated to gross findings. The third group shows nearly complete regeneration of appendage structure similar to normal in the dermis in contrast to control and second group with absence and less number of skin appendages, respectively.