Procollagen Synthesis Research Articles

Effects of Bletillae Rhizoma on the Elastase, Collagenase, and Tyrosinase Activities and the Procollagen Synthesis in Hs68 Human Fibroblasts

Objectives： Bletillae Rhizoma, the roots of Bletilla striata, is used to restrain the leakage of blood and stop bleeding. It can cure the sores, ulcers, and chapped skin. This study was designed to investigate the collagen metabolism, elastase and tyrosinase activity of Bletillae Rhizoma extract (BR). Methods : The effects of BR on type I procollagen production and collagenase activity in human normal fibroblasts Hs68 after UVB (312 nm) irradiation were measured by ELISA method. The elastase activity, tyrosinase activity, and L-DOPA oxidation after treatment of BR were measured as well. Results : In the present study, the collagen production (type I procollagen) was significantly increased to <TEX>$15.7{\pm}1.8$</TEX> ng/ml at a concentration of BR 100 <TEX>${\mu}g/ml$</TEX> in UVB damaged Hs68 cells. The increased collagenase activity after UVB damage was significantly recovered to <TEX>$42.7{\pm}0.7%$</TEX>, <TEX>$54.5{\pm}3.5%$</TEX>, and <TEX>$38.4{\pm}0.9%$</TEX> by BR 10, 30, and 100 <TEX>${\mu}g/ml$</TEX>. The activities of BR 10 mg/ml on tyrosinase activity was significantly reduced to <TEX>$45.1{\pm}8.4%$</TEX> as well. However, there were no significant effects on the elastase activity and the L-DOPA oxidation. Conclusion : BR showed the promoting effects of collagen synthesis and inhibitory effects of collagenase activity in Hs68, human normal fibroblast cells. And these could be thought to have the anti-wrinkle effects and whitening effects in vitro. These results suggest that BR may have potential as an anti-aging ingredient in cosmetic treatment.

The effect of cilostazol on the expression of matrix metalloproteinase-1 and type I procollagen in ultraviolet-irradiated human dermal fibroblasts

AimCilostazol is a selective inhibitor of type III phosphodiesterase that inhibits platelet aggregation. Cilostazol is a useful vasodilator, antithrombotic, and cardiotonic agent. Ultraviolet B (UVB) irradiation increases the production of matrix metalloproteinase-1 (MMP-1) during skin photoaging. The UVB-induced increase of MMP-1 results in connective tissue damage, and the skin becomes wrinkled and aged. Here, we investigated the capacity of cilostazol to inhibit MMP-1 expression in UVB-irradiated human dermal fibroblasts. Main methodsCultured human dermal fibroblasts were irradiated with UVB, followed by the addition of cilostazol to the culture medium. Key findingsPost-treatment with cilostazol attenuated UVB-induced production of MMP-1 and prevented the reduction of type I procollagen. Cilostazol inhibited UVB irradiation-induced phosphorylation of the mitogen-activated protein kinase (MAPK) signaling molecules Jun-N-terminal kinase (JNK) and p38 kinase, as well as activator protein-1 (AP-1) in dermal fibroblasts. SignificanceOverall, these results demonstrate that cilostazol regulates UVB-induced MMP-1 expression and type I procollagen synthesis by inhibiting MAPK signaling and AP-1 activity. Therefore, we suggest that cilostazol may be useful for the prevention and treatment of skin photodamage caused by UVB-irradiation.

양마에서 분리한 Kaempferol과 그 배당체의 항균 및 주름억제 효과

In this study, kaempferol and its rhamnosides (α-rhamnoisorobin, afzelin, kaempferitrin) were isolated electively by bioconversion technology from Hibiscus cannabinus L. leaves to evaluate the anti-wrinkles effects and anti-microbial effects. In order to evaluate anti-wrinkles activity, reduction of expression matrix metalloproteinase-1 (MMP-1) protein and proliferation/ pro-collagen production were investigated. Kaempferol and α-rhamnoisorobin showed inhibition activity of MMP-1 generated to compared to positive control. In HaCaT cell proliferation assay, kaempferol and α-rhamnoisorobin significantly promoted cell proliferation in a concentration-dependent manner. In addition, procollagen synthesis assays (by HDF-N cell) showd that TGF-β induced procollagen production and also, all four kinds of experimental significantly promoted procollagen synthesis in a concentration-dependent manner. Kaempferol and α-rhamnoisorobin exhibited strong antimicrobial activities on five of microbes, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger.

Ascorbic acid enhances the expression of type 1 and type 4 collagen and SVCT2 in cultured human skin fibroblasts

Ascorbic acid (AA) is essential for collagen biosynthesis as a cofactor for prolyl and lysyl hydroxylase and as a stimulus for collagen gene expression. Many studies have evaluated the relationship between AA and collagen expression in short- and long-term effects on cells after a single administration of AA into the culture medium. However, no such study has monitored in detail the stability of AA in medium or the alterations of intracellular AA levels during a protracted interval. Therefore, we examined here intracellular AA levels and stability throughout its exposure to human skin fibroblasts in vitro. Moreover, we determined the effects on type 1 and type 4 collagen and sodium-dependent vitamin C transporter (SVCT) gene expression when medium containing 100μM AA was replaced every 24h for 5days to avoid depletion of AA. Throughout this long-term culture, intracellular AA levels remained constant; the expression of type 1 and type 4 collagens and SVCT2 mRNA was enhanced, and type 1 procollagen synthesis increased. Thus, these results indicate that human skin fibroblasts exposed to AA over time had rising levels of type 1/type 4 collagens and SVCT2 mRNA expression and type 1 procollagen synthesis.

Efficacy of a Novel Treatment Serum in the Improvement of Photodamaged Skin

SynopsisA novel treatment serum formulated to target multiple pathways in the anti-ageing cascade was tested both in vitro and in clinical settings. In vitro testing was performed to assess the ability to stimulate key proteins and genes fundamental to the anti-ageing cascade. The antioxidant potential of the formulation was studied in a UV-irradiation clinical study. A 12-week, open-label, single-centre study was conducted to determine whether this uniquely formulated topical treatment serum could improve visible signs of facial photodamage. Clinical evaluations showed statistically significant reductions in fine wrinkles and coarse wrinkles and improvements in skin texture, tone and radiance starting at week 4 with continued improvements at weeks 8 and 12. Subject self-assessments confirmed that the beneficial effects of the treatment serum were readily observed by the users. The treatment serum was well tolerated with no treatment-related adverse events reported during the 12-week study. Use of this novel treatment serum produced significant improvements in the visible signs of facial photodamage.RésuméUn nouveau sérum de traitement conçu pour cibler de multiples voies dans la cascade anti-âge a été testé à la fois in vitro et dans les conditions cliniques. Les tests in vitro ont été réalisés afin d'évaluer la capacité de stimuler les protéines et les gènes clés fondamentales de la cascade anti-vieillissement. Le potentiel antioxydant de la formulation a été étudié dans une étude clinique utilisant le rayonnement UV. Une étude de douze semaines, en mode ouvert, monocentrique a été menée afin de déterminer si ce sérum spécialement formulé pour le traitement topique peut améliorer les signes visibles du photo-vieillissement du visage. Des évaluations cliniques ont montré une réduction statistiquement significative des rides et des ridules secondaires et l'amélioration de la texture de la peau, du tonus et d'éclat à partir de la semaine 4 avec des améliorations continues aux semaines 8 et 12. Les autoévaluations par les sujets ont confirmé que les effets bénéfiques du sérum de traitement étaient facilement observés par les utilisateurs. Le sérum de traitement a été bien toléré avec aucun événement indésirable rapporté au cours de l'étude de 12 semaines. L'utilisation de ce nouveau sérum de traitement produit des améliorations significatives dans les signes visibles du photovieillissement du visage.

Fibrosis and renin-angiotensin-aldosterone system activity. Reality and future prospects

Myocardial fibrosis plays a key role in the pathogenesis of cardiovascular diseases. Renin-angiotensin-aldosterone system (RAAS) is the key regulator of vascular tone, potassium and water homeostasis, and response to the tissue damage. The effects of angiotesin II are mediated by 1 type angiotensin II receptors, together with angiotensin-converting enzyme, forming the «classical regulation axis» of RAAS. At chronic high cardiac afterload the genes of procollagen and collagen synthesis are overexpressed leading to the increased production of collagen, fibrosis and myocardial hypertrophy. Both hemodynamic and non-hemodynamic factors affect fibrosis development. RAAS blockers are suggested to have antifibrotic activity. There is a strong evidence of losartan efficiency that can be, at least, partly mediated by antifibrotic activity.

우방자에서 분리한 주름개선 화합물

This study was carried out to discover the skin wrinkle reducing components in the seeds of Arctium lappa. The isolation of a methylene chloride-soluble fraction of 70% ethanol extract from the seeds of Arctium lappa using a procollagen type-1 synthesis and MMP-1 activity resulted in the isolation and identification of four lignin compounds: arctiin, arctigenin, matairesinol, and diarctigenin. All structures were confirmed via NMR and MS spectroscopic data. To determine cell viability and procollagen type-1 synthesis, human dermal fibroblasts were treated with 10-100 μM. As a result, none of the four compounds showed cytotoxicity up to 50 μM. We also investigated their procollagen type-1 synthesis and MMP-1 inhibition activity and found that arctiin had the highest activity in terms of both procollagen synthesis and MMP-1 inhibition among all four compounds. Putting all the data together, we suggest that arctiin be used in cosmetics as an anti-wrinkle material.

Inhibitory effect of Astragalus membranaceus root on matrix metalloproteinase-1 collagenase expression and procollagen destruction in ultraviolet B-irradiated human dermal fibroblasts by suppressing nuclear factor kappa-B activity

The root of Astragalus membranaceus, regarded as a tonic in traditional Korean medicine, has been prescribed for long periods to treat chronic illness by boosting the immune system. Ultraviolet (UV) irradiation causes damage to skin connective tissue by degrading collagen, which is a major structural component of the extracellular matrix. Such damage is considered to be a cause of the wrinkling observed in premature ageing of the skin. This study has investigated the photo-protective effect of A. membranaceus on UVB radiation-induced activation of nuclear factor kappa-B (NF-κB) activity in human dermal fibroblasts. HS68 fibroblast cells cultured with various concentrations of A. membranaceus were exposed to UVB (40 mJ/cm²). Activation of NF-κB P65 and expression of matrix metalloproteinase-1 (MMP-1) and type 1 procollagen were measured by Western blotting. Translocation of NF-κB P65 and MMP-1 regulation were also examined by immunocytochemistry. Western blotting and immunocytochemistry results showed that A. membranaceus inhibited UVB-induced translocation of NF-κB P65 and MMP-1 expression. The data suggested that A. membranaceus restored type 1 procollagen synthesis by inhibiting NF-κB P65 activity and MMP-1 expression in UVB-exposed human dermal fibroblasts. A. membranaceus is a candidate for use in skin protection from UVB-induced skin inflammation and photoageing.

Abstract 208: Farnesylation-Dependent Fibrosis in the Aged Murine Heart

Mesenchymal stem cells (MSC) derived from the aged murine heart express reduced levels of the multipotency marker Nanog. These CD44 + CD45 - MSC inappropriately differentiate into fibroblasts characterized by markedly enhanced basal collagen expression. While collagen synthesis in cardiac fibroblasts derived from young mice is driven by TGF-β1, paradoxically, fibroblasts derived from the aged heart displayed reduced TGF-β1 responsiveness and decreased TGF-β receptor expression (ALK5). To resolve this apparent contradiction we looked for other pathways leading to collagen expression. We found that circulating insulin levels in aged mice were elevated ∼2.5 fold when compared to young animals. We hypothesized that increased circulating insulin levels drive MSC differentiation and caused upregulation of procollagen expression in cardiac fibroblasts. Our in vitro experiment confirmed that insulin at a pathophysiological concentration (1 nM) increased procollagen type I expression only in fibroblasts isolated from old hearts. As insulin can signal through substrate prenylation, cardiac fibroblasts isolated from aged mice had ∼2 fold greater basal activity of farnesyltransferase (FTase, enzyme that transfers the prenyl chain) than fibroblasts isolated from young mice. FTase activity was further increased when fibroblasts isolated from aged hearts were stimulated with insulin. Cells derived from 3 month old mice did not upregulate FTase activity in response to insulin. Basal expression of FTase regulatory subunit (FNTA) was also upregulated by 3 fold in aged fibroblasts and expression of catalytic subunit (FTNB) was increased with insulin stimulation. FTase inhibitor reduced procollagen synthesis in aged fibroblasts. Here we present new data linking insulin-dependent upregulation of collagen type I synthesis via increased farnesylation in the aging heart.

BBF2H7-Mediated Sec23A Pathway Is Required for Endoplasmic Reticulum-to-Golgi Trafficking in Dermal Fibroblasts to Promote Collagen Synthesis

Collagen fibers, structural elements responsible for mechanical strength in skin, are synthesized constitutively in response to cytokines such as IGF-I. However, little is known about their intracellular trafficking from the endoplasmic reticulum (ER) to the Golgi apparatus during synthesis. We demonstrate herein that the BBF2 human homolog on chromosome 7 (BBF2H7)-mediated Sec23A pathway is involved in regulation of intracellular procollagen trafficking. The mRNA and protein expression of BBF2H7, Sec23A, and type I and III collagen (COL1 and COL3) was induced by IGF-I stimulation. In addition, the cleaved form of BBF2H7 was detected in IGF-I-treated cultures, indicating that activation occurs concurrently with its expression. Knockdown with small interfering RNAs targeting BBF2H7 caused a significant reduction in the expression of COL1 and COL3, regardless of IGF-I treatment. Both mitogen-activated protein kinase and phosphatidylinositol-3 kinase pathways via IGF-I receptor activation were required for BBF2H7 induction. Using immunofluorescence microscopy, we showed that Golgi apparatus dysmorphology is due to coat protein complex II vehicle hypoplasia caused by the absence of BBF2H7 and Sec23A. The BBF2H7-mediated Sec23A pathway was required for ER-to-Golgi procollagen trafficking to promote collagen synthesis. This role of growth factors such as IGF-I, which to our knowledge is previously unreported, suggests antiaging strategies.

Anti‐ageing effects of alpha‐naphthoflavone on normal and UVB‐irradiated human skin fibroblasts

Ageing is a complex and multifactorial process resulting in several functional and aesthetic changes to the skin. We found that α-Naphthoflavone (α-NF) concentration-dependently induced pro-collagen type I protein expression and inhibited MMP-1 protein expression, in both normal and UVB-irradiated cells. SB431542 and SIS3 - inhibitors of TGF-β and Smad3, respectively - significantly alleviate α-NF-caused response of MMP-1 and pro-collagen. LY294002 (PI3K inhibitor) can reverse α-NF-induced ERK, Akt, Smad-3 activation, pro-collagen synthesis and α-NF-suppressed AP-1 activation. PD (ERK inhibitor) was not involved in pro-collagen generation and MMP-1 inhibition. We concluded that α-NF promotes pro-collagen production and inhibits MMP-1 expression via the activation of a PI3K/Akt/Smad-3 pathway in normal and UVB-irradiated human skin fibroblasts, while TGF-β may play an important role in transducing this pathway. These results suggest that α-NF, a natural plant product, has the potential to become a novel anti-ageing skin application.

Inhibitory effects of sea buckthorn (Hippophae rhamnoides L.) seed on UVB-induced Photoaging in human dermal fibroblasts

Sea buckthorn (Hippophae rhamnoides L.) seed, which has very high nutraceutical, cosmetoceutical and therapeutic properties, has been widely used to treat diseases in Tibetan and Mongolian traditional medicines. In this study, we measured the antioxidant activities of the solvent-dependent SBS (Sea buckthorn seed) extracts using the DPPH and Folin-Ciocalteu assays and we investigated the efficacy of SBS in protecting skin against UVB-induced damage using cultured human dermal fibroblasts. In addition, the inhibitory effects of SBS on UVB-induced skin photoaging were examined by determining the level of Metalloproteinase (MMP)-1 and procollagen after UVB-irradiation. The antioxidant capacity of the SBSE (alcoholic) extract was significantly higher than the SBSH (hexane) and SBSW (water) extracts, as measured based on the free-radical scavenging activity and total phenolic content. SBSE was chosen as the most suitable candidate antioxidant. In human dermal fibroblasts, the cell viability of SBSE extract at doses of 2.5, 5, and 10 μg/mL was higher than the UVB-treated control. By RT-PCR and Western blot, SBSE treatment inhibited UVB-induced IL-1β expression in cultured cells. In addition, SBSE restrained UVB-induced IL-6 and COX-2 gene expression in cultured fibroblasts in a dose-dependent manner. Treatment with the SBSE extract after UVB irradiation in human dermal fibroblasts significantly reduced MMP-1 expression and increased procollagen synthesis when compared with UVB-irradiation only. In this study, SBSE was shown to increase the synthesis of procollagen, decrease the expression of MMP-1, and inhibit the production of IL-1β, IL-6, and COX-2 in UVB-irradiated human fibroblasts. These findings suggest that the SBSE extract may be a potential therapeutic agent for preventing and treating skin photoaging.

Dioscorea batatas stimulates procollagen production in vitro and reduces UVB‐induced MMPs secretion in vivo

Ultraviolet (UV) B irradiation induces the production of matrix metalloproteinases (MMPs), which are responsible for the degradation or synthesis inhibition of collagenous extracellular matrix in connective tissues, causing skin photoaging. In this study, we examined the efficacy of a yam extract in protecting skin against UVB‐induced damage using cultured human dermal fibroblast neonatal (HDFn) cells and hairless mice skin. Dioscorea batatas (DB) was used the synthesis of procollagen and the release of MMP‐1 in HDFn cells were measured by EIA kit and MMP‐1 assay kit, respectively. UVB radiation was applied to the backs of the mice three times a week for 8 weeks. Mice were randomly divided into four groups, and were topical application with the DB (3% and 6%) or vehicle. DB (50 ug/ml) stimulated procollagen production in HDFn cells by 482.4 ng/ml compared to TNF‐á– treated cell (269.5 ng/ml). Also, pre‐treatment of HDFn cells with DB inhibited TNF‐á–induced release of MMP‐1. In vivo study, we found that DB attenuated UVB‐induced epidermal thickness. DB suppressed the expression of MMP‐3 and MMP‐13 induced by UVB irradiation. Our results show that the DB protects against UV‐induced skin damage in hairless mice. This work was supported by the Ministry of Knowledge Economy, Republic of Korea (70007085).

Effects of matrix metallopeptidase 9 on the proliferation, apoptosis, type I and III procollagen synthesis of rat dermal fibroblasts

To explore the effects of matrix metallopeptidase 9 (MMP-9) on the proliferation, apoptosis, type I and III procollagen synthesis of rat dermal fibroblasts. Lipopolysaccharide (LPS) at 0.1 and 1.0 µg/ml was used to stimulate fibroblasts to up-regulate the expression of MMP-9 for 18 and 48 h. And then RNA interference was employed to inhibit the high expression of MMP-9. Cell proliferation was tested by CCK-8, cell apoptosis by flow cytometry and type I and III procollagen expressions by quantitative reverse transcriptase PCR (qRT-PCR). The MMP-9 expression of fibroblasts increased after the stimulation of LPS. And the 1.0 µg/ml LPS stimulation of 48 hours was 14.25 times of mRNA expression and 2.31 times of protein expression versus that of the normal group (both P < 0.05). The RNA interference obviously inhibited the high expression of MMP-9. The mRNA expression was 1/8 and protein expression 1/3 (both P < 0.05) as compared with the control group. Cell proliferation decreased with the rising expression of MMP-9 to some extent [(1.08 ± 0.08) vs (1.18 ± 0.09), P < 0.05] and improved after the inhibition of high expression of MMP-9 [(1.78 ± 0.17) vs (1.53 ± 0.15), P < 0.01]. There was no change of apoptosis accompanied with high expression of MMP-9 (P > 0.05). Apoptosis decreased after the inhibition of high expression of MMP-9 for 48 hours (3.53% ± 0.22% vs 4.47% ± 0.46%, P < 0.05). The synthesis of type I procollagen was the same no matter up-regulation or down-regulation of MMP-9 expression (P > 0.05). As the expression of MMP-9 increased, the synthesis of type III procollagen decreased (P < 0.01), but not increased by 1.02 folds after the inhibited expression of MMP-9 (P > 0.05). MMP-9 can affect the biological behaviors of rat dermal fibroblasts.

Fibrodysplasia Induced in Dog Skin by a Matrix Metalloproteinase (MMP) Inhibitor—A Mechanistic Analysis

Matrix metalloproteinase (MMP) inhibitors, candidate therapeutic agents for a number of diseases, are known to be associated with acute fibrosis-type adverse effects in a number of species, including humans. The broad-spectrum MMP inhibitor, AZM551248, has previously been shown to cause these effects in the dog. Changes were characterized by the abnormal and extensive proliferation of fibroblasts and the deposition of collagen particularly in the subcutaneous connective tissues (subcutis) and were termed fibrodysplasia (FD). We performed a time-course study in dogs using AZM551248 and sampled skin, subcutis, and plasma before and during the development of FD. Detailed histopathological analysis and global gene expression profiling were performed on the subcutaneous tissues. The gene expression analysis of the subcutis indicated that extracellular matrix (ECM) remodeling was initiated asymptomatically at or before the earliest time point, day 4, and this was associated with dysregulation of expression of a number of MMPs and proteolytic enzymes. At later time points, the FD became progressively more extensive and severe, and this was associated with gene expression changes characteristic of tissue fibrosis, for example those associated with procollagen synthesis and processing. We postulate that AZM551248 inhibition of MMP action within the subcutis modulates the activity of several transcription factors and this in turn upregulates expression of specific proteases which initiate ECM remodeling. Persistent MMP inhibition results in the progression of ECM remodeling, culminating in collagen deposition and overt fibrosis. Our data indicate that inhibition of MMPs 1, 2, 3, and 9 is a key early event in AZM551248-induced FD in dog subcutis.

Femoral Artery Dissection in Vascular Type Ehlers–Danlos Syndrome; Leave Well Alone?

Vascular Ehlers–Danlos Syndrome (EDS) is a rare autosomal dominant condition resulting from a defect in type III procollagen synthesis. This causes the development of severe vascular pathologies, including arterial rupture and pseudoaneurysm formation. We present a case of a young boy previously diagnosed with vascular EDS due to a Gly975Val substitution in the collagen α1(III) chain presenting with a common femoral artery dissection secondary to minimal trauma. This was managed conservatively with serial duplex scans and gentle mobilization. At follow up the patient had returned to normal activities, with MRA and duplex scans showing complete resolution of the dissection.

The retinoic acid-induced up-regulation of insulin-like growth factor 1 and 2 is associated with prolidase-dependent collagen synthesis in UVA-irradiated human dermal equivalents

Ultraviolet (UV) A irradiation causes the degeneration of extracellular matrix in the skin dermis, mainly due to disrupted collagen homeostasis, resulting in the photo-aging of human skin. All-trans retinoic acid (ATRA) improves photo-aged human skin in vivo. Although the effects of ATRA on collagen synthesis and MMP regulation are well known, the effects of ATRA on other collagen homeostasis-associated genes have not been elucidated. This study was aimed to study the factors that are pharmacologically associated with the effect of ATRA on collagen homeostasis. The gene transcription profile of collagen homeostasis-associated genes was systematically evaluated in three-dimensional human dermal equivalents (HDEs) following UVA-irradiation and/or ATRA treatment. In addition to the expected changes in MMPs and collagen synthesis in HDEs in response to ATRA, prolidase, an important enzyme in the recycling of proline and hydroxyproline from degraded collagen molecules, was significantly decreased by UVA irradiation, and its down-regulation was antagonized by ATRA. Transfection with a prolidase-specific siRNA led to a significant decrease in procollagen synthesis in human fibroblasts. ATRA inhibited the UVA irradiation-induced decrease in prolidase activity through an insulin-like growth factor (IGF) receptor signaling pathway in HDEs. ARTA increased IGF1 and IGF2 production in HDEs, and neutralizing IGFs with anti-IGF antibodies abolished the effect of ATRA on proliase activity. These data demonstrate that ATRA regulates prolidase activity in HDEs via IGF receptor signaling, suggesting one of the pharmacological mechanisms by which improves photo-aged human skin.

Dermal penetration of creatine from a face-care formulation containing creatine, guarana and glycerol is linked to effective antiwrinkle and antisagging efficacy in male subjects

The dermal extracellular matrix provides stability and structure to the skin. With increasing age, however, its major component collagen is subject to degeneration, resulting in a gradual decline in skin elasticity and progression of wrinkle formation. Previous studies suggest that the reduction in cellular energy contributes to the diminished synthesis of cutaneous collagen during aging. To investigate the potential of topically applied creatine to improve the clinical signs of skin aging by stimulating dermal collagen synthesis in vitro and in vivo. Penetration experiments were performed with a pig skin ex vivo model. Effects of creatine on dermal collagen gene expression and procollagen synthesis were studied in vitro using cultured fibroblast-populated collagen gels. In a single-center, controlled study, 43 male Caucasians applied a face-care formulation containing creatine, guarana extract, and glycerol to determine its influence on facial topometric features. Cultured human dermal fibroblasts supplemented with creatine displayed a stimulation of collagen synthesis relative to untreated control cells both on the gene expression and at the protein level. In skin penetration experiments, topically applied creatine rapidly reached the dermis. In addition, topical in vivo application of a creatine-containing formulation for 6 weeks significantly reduced the sagging cheek intensity in the jowl area as compared to baseline. This result was confirmed by clinical live scoring, which also demonstrated a significant reduction in crow's feet wrinkles and wrinkles under the eyes. In summary, creatine represents a beneficial active ingredient for topical use in the prevention and treatment of human skin aging.

Ixora parviflora Protects against UVB-Induced Photoaging by Inhibiting the Expression of MMPs, MAP Kinases, and COX-2 and by Promoting Type I Procollagen Synthesis.

Ixora parviflora with high polyphenol content exhibited antioxidant activity and reducing UVB-induced intracellular reactive oxygen species production. In this study, results of the photoaging screening experiments revealed that IPE at 1000 μg/mL reduced the activity of bacterial collagenase by 92.7 ± 4.2% and reduced the activity of elastase by 32.6 ± 1.4%. Therefore, we investigated the mechanisms by which IPE exerts its anti-photoaging activity. IPE at 1 μg/mL led to an increase in type I procollagen expression and increased total collagen synthesis in fibroblasts at 5 μg/mL. We found that IPE inhibited MMP-1, MMP-3, and MMP-9 expression at doses of 1, 5, and 10 μg/mL, respectively, in fibroblasts exposed to UV irradiation (40 mJ/cm2). Gelatin zymography assay showed that IPE at 50 μg/mL inhibited MMP-9 secretion/activity in cultured fibroblasts after UVB exposure. In addition, IPE inhibited the phosphorylation of p38, ERK, and JNK induced by UVB. Furthermore, IPE inhibited the UVB-induced expression of Smad7. In addition, IPE at 1 μg/mL inhibited NO production and COX-2 expression in UV-exposed fibroblasts. These findings show that IPE exhibits anti-inflammatory and anti-photoaging activities, indicating that IPE could be a potential anti-aging agent.

Seeding a human tendon matrix with bone marrow aspirates compared to previously isolated hBMSCs – An in vitro study

Injuries of tendons and ligaments give rise to significant morbidity. Tissue engineering offers promising treatment concepts such as seeding a scaffold with human bone marrow stem cells (hBMSCs) to create high-quality tendon replacement tissue. HBMSCs are usually isolated and cultured prior to seeding. Studies evaluating if previous isolation is superior to seeding with bone marrow aspirates have not been published yet. The aim of this study was to compare these two seeding methods in terms of cell viability, proliferation and differentiation. HBMSCs were harvested from the iliac crest during routine trauma surgery. In group A the scaffold (human achilles tendons) was seeded with bone marrow aspirates, while in group B hBMSCs were isolated, harvested and then seeded. Constructs were stimulated in perfusion bioreactors according to established protocols. Mean cell proliferation was significantly increased (p< 0.05) on tendons seeded with bone marrow aspirates. Cell viability, the concentration of alkaline phosphatase in the perfused media and the synthesis of procollagen - III - polypeptide (PIIIP) were not significantly different when comparing the two groups. The synthesis of procollagen-I-polypeptide (PIP) was significantly increased on tendons seeded with previously isolated hBMSCs p < 0.05). The results indicate that seeding a human tendon matrix scaffold with bone marrow aspirates may be equal to seeding with previously isolated hBMSCs. This new seeding method could facilitate and speed up the tissue engineering process.