The Combined Effect of Lumenato and Ceramide in the Protection of Collagen Damage Induced by Neutrophils in Normal Human Dermal Fibroblasts

Introduction: Collagen is the primary structural protein fibroblasts produce in the skin’s extracellular matrix. Infiltration of neutrophils into the epidermis and dermis by exposure to UV causes collagen damage and contributes to photoaging. Methods: To study the combined effect of Lumenato and ceramide in preventing collagen-1 damage induced by phagocytes, we used co-cultures of normal human dermal fibroblasts (fibroblasts) and activated human neutrophils. The present study aimed to determine the protective effect of the combination of Lumenato and ceramide on fibroblast collagen-1 damage induced by neutrophils. Results: Lumenato (in the range of 6.5 - 208 μg/ml) or ceramide (in the range of 0.1 - 50 μM) inhibited the production of superoxides and MPO by TNFα-stimulated neutrophils, as well as the production of NO by LPS-stimulated macrophages in a dose-dependent manner. The combinations of Lumenato and ceramide, in low concentrations, caused synergistic prevention of fibroblasts’ collagen-1 damage induced by TNFα-activated neutrophils, detected by fluorescence immunostaining and WB analysis. MPO activity in the supernatants of the co-cultures was also synergistically inhibited. Adding Lumenato or ceramide singly or in combinations in these low concentrations to the fibroblast cultures did not affect the expression of collagen-1. The combinations of Lumenato or ceramide in these concentrations also caused a synergistic inhibition of NO production by activated macrophages. Conclusions: The results suggest that combining low concentrations of Lumenato and ceramide results in synergistic protection against fibroblasts’ collagen-1 damage induced by neutrophils, thus indicating their possible potential for enhanced skin health.

Chicken collagen hydrolysates differentially mediate anti-inflammatory activity and type I collagen synthesis on human dermal fibroblasts

Collagen is a major extracellular matrix protein.Given the potential anti-inflammatory and antioxidant profiles of these bioactive compounds,there has been increasing interest in using collagen derived peptides and peptide-rich collagen hydrolysates for skin health,due to their immunomodulatory,antioxidant and proliferative effects on dermal fibroblasts.However,all hydrolysates are not equally effective in exerting the beneficial effects;hence,further research is needed to determine the factors that improve the therapeutic applicability of such preparations.We used different enzymatic conditions to generate a number of different collagen hydrolysates with distinct peptide profiles.We found that the use of two rather than one enzyme for hydrolysis generates a greater abundance of low molecular weight peptides with consequent improvement in bioactive properties.Testing these hydrolysates on human dermal fibroblasts showed distinct actions on inflammatory changes,oxidative stress,type I collagen synthesis and cellular proliferation.Our findings suggest that different enzymatic conditions affect the peptide profile of hydrolysates and differentially regulate their biological activities and potential protective responses on dermal fibroblasts.

The Polypeptide in Chlamys farreri can protect human dermal fibroblasts from ultraviolet B damage

To investigate the effect of polypeptide from Chlamys farreri (PCF) on NHDF in vitro, we modeled oxidative damage on normal human dermal fibroblasts (NHDF) exposed to ultraviolet B (UVB). In this study, 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydro-genase (LDH) were tested to measure cell viability. Enzymes including superoxide dismutase (SOD), glu-tathione peroxidase (GSH-PX), catalase (CAT) and xanthine oxidase (XOD) were determined biochemically. Total antioxidative capacity (T-AOC) and anti-superoxide anion capacity (A-SAC) were also determined. Ultrastructure of fibroblasts was observed under transmission electron microscope. The results showed that: UVB (1.176×10-4 J/cm2) suppressed the growth of fibroblasts and the introduction of PCF (0.25%-l%) before UVB reduced the suppression in a concentration-dependent manner. PCF could enhance the activities of SOD, GSH-PX and T-AOC as well as A-SAC. Also PCF could inhibit XOD activity, while it did not affect CAT activity. Ultrastructure of fibroblasts were damaged after UVB irradiation, concentration-dependent PCF reduced the destructive effect of UVB on cells. These results indicated that PCF can protect human dermal fibroblasts from being harmed by UVB irradiation via its antioxidant pro-erty.

9-Hydroxy-6,7-dimethoxydalbergiquinol suppresses hydrogen peroxide-induced senescence in human dermal fibroblasts through induction of sirtuin-1 expression

Objective:To investigate the potential anti-aging mechanism of9-hydroxy-6,7-dimethoxydalbergiquinol(HDDQ)on hydrogen peroxide(H2O2)-induced oxidative stress in human dermal fibroblasts(HDFs).Methods:The effect of HDDQ on cell viability was assessed by MTT assay,and the effects of HDDQ on senescence-like phenotypes were determined by senescence-associatedβ-galactosidase(SA-β-gal)staining,Western blotting analysis,and a cell proliferation assay.The expression level and activity of sirtuin-1(SIRT1)induced by HDDQ were also measured.Results:HDDQ reversed senescence-like phenotypes in the oxidant-challenged model,through reducing SA-β-gal activity and promoting cell growth.Meanwhile,decreases in ac-p53,p21Cip1/WAF1,and p16Ink4a and an increase in p Rb were observed.HDDQ induced the expression of SIRT1 in a concentration-and time-dependent manner.Moreover,HDDQ inhibited H2O2-induced phosphorylation of Akt by SIRT1 up-regulation and reduced SA-β-gal staining.Conclusions:HDDQ inhibits H2O2-induced premature senescence and upregulation of SIRT1 expression plays a vital role in the inhibition of the senescence phenotype in HDFs.

Anti-senescence and anti-wrinkle activities of 3-bromo-4,5-dihydroxybenzaldehyde from Polysiphonia morrowii Harvey in human dermal fibroblasts

Objective:To investigate the anti-senescence effect of 3-bromo-4,5-dihydroxybenzaldehyde(BDB)from Polysiphonia morrowii Harvey in human dermal fibroblasts(HDF).Methods:HDF were subjected to treatment of BDB and then treated with hydrogen peroxide(H2O2)to induce premature senescence.Senescence-associatedβ-galactosidase(SA-β-gal)activity in HDF was determined using the SA-β-gal staining method.Intracellular reactive oxygen species(ROS)production was measured using the 2’,7’-dichlorodihydrofluorescein diacetate assay.Western blotting assay was performed to assess the level of antioxidant enzyme glutathione peroxidase 1(GPX1).In addition,intracellular collagen and collagenase contents were analyzed using the respective ELISA kits.Elastase activity in HDF supernatants was measured from p-nitroaniline release and normalized using total protein content.Results:Treatment of HDF with H2O2 increased the activity of SA-β-gal,but BDB pre-treatment resulted in the reduction of SA-β-gal activity.Moreover,BDB significantly reduced H2O2-induced intracellular ROS production.BDB also markedly increased the level of GPX1,which was inhibited by 400μM of H2O2.Furthermore,in in vitro study,BDB significantly increased intracellular collagen content and decreased matrix metalloproteinase-1 and elastase activities in HDF.Conclusions:Our results demonstrate that BDB shows antisenescence and anti-wrinkle activities in vitro.

Effect of a Nutrient Mixture on Fanconi Anemia Fibroblast and Normal Human Dermal Fibroblast: A Comparison

Fanconi anemia (FA) is a fatal heterogeneous autosomal recessive disorder, characterized by progressive bone marrow failure, congenital defect and cancer predisposition. Cell culture from FA fibroblast (FAF) displays certain abnormalities as compared to normal human dermal fibroblast (NHDF). This prompted us to investigate the effect of a specific nutrient mixture (NM) containing ascorbic acid, lysine, proline and green tea extract, which has demonstrated a broad spectrum of pharmacological activities, on FAF compared to NHDF. We investigated the in vitro effect of NM on FAF and NHDF cell proliferation by MTT assay, MMPs secretion by zymography, morphology by H&E staining and apoptosis by green caspase assay. FAF (FA-A: PD20, FA-A: PD220) and NHDF were cultured in modified Dulbecco Eagle media. At near confluence, the cells were treated with different concentrations of NM (0, 50, 100, 250, 500 and 1000 μg/ml) in triplicate. The cells were also treated with PMA to induce MMP-9 activity. NM had no effect on FAF cell viability in both cell lines compared to control. In contrast NM exhibited 20% at 50 and 100, 50% at 250, 60% at 500 and 70% toxicity at 1000 μg/ml on NHDF cells. Zymography demonstrated MMP-2 and MMP-9 on PMA stimulation in FAF and NM inhibited the activity of both MMP-2 and MMP-9 in a dose response fashion with total block at 500 μg/ml. In contrast, NHDF exhibited only MMP-2, both active and inactive forms, and NM inhibited their activities in a dose-dependent manner with total block at 1000 μg/ml. H&E staining did not indicate any morphological changes in FAF nor induced apoptosis at higher concentrations, as seen by caspases assay. However, although no morphological changes in NHDF were noted up to NM 100 μg/ml, progressive changes in cell shrinkage, rounding and nuclear condensation, pertaining to apoptosis, were observed at higher concentrations. These changes were consistent with the results from the green caspases apoptosis assay. Our data demonstrate that NM exhibited different responses toward FAF and NHDF. This may in part be due to elevated chromosomal break, deletion and hypersensitivity to cross linking agents, a DNA repair disorder in FAF that is lacking in NHDF.

EXPERIMENTAL STUDY ON INDUCTION OF OSTEOGENETIC POTENTIAL OF DERMAL FIBROBLASTS

Objectiye To eiucidate the inducing factor in osteogenesis piayed by dermai fibroblasts. mthods Fibroblasts and epithelial cells were procured from rabbit and human skins and separated. These two types of cells were then cultured either together or separately and studied histochemically, immunohistochemically and biochemically. Results Bone nodules were found only in the mtxed culture of fibroblasts and epithelial cells. During the period ol exuberant bone nodule formation, there were both more positive keratin- stained cells (epithelial cells) and higher ALP activity. When few or no bone nodules formed, positive keratin- stained cells declined or disappeared and the ALP activity fell off. Conclusion That dermal fibroblasts show their osteogenic potential depends mainly on the inducing effect of the ePithelial cells.

Dermal fibroblast expression of stromal cellderived factor-1 （SDF-1） promotes epidermal keratinocyte proliferation in normal and diseased skin

Stromal cells provide a crucial microenvironment for overlying epithelium. Here we investigated the expression and function of a stromal cell-specific protein, stromal cell-derived factor-1 （SDF-1）, in normal human skin and in the tissues of diseased skin. Immunohistology and laser capture microdissection （LCM）-coupled quantitative real- time RT-PCR revealed that SDF-1 is constitutively and predominantly expressed in dermal stromal cells in nor- mal human skin in vivo. To our surprise, an extremely high level of SDF-1 transcription was observed in the dermis of normal human skin in vivo, evidenced by much higher mRNA expression level than type I collagen, the most abundant and highly expressed protein in human skin. SDF-1 was also upregulated in the tissues of many human skin disorders including psoriasis, basal cell carcinoma （BCC）, and squamous cell carcinoma （SCC）. Double immunostaining for SDF-1 and HSP47 （heat shock protein 47）, a marker of fibroblasts, revealed that fibroblasts were the major source of stroma-cell-derived SDF-1 in both normal and diseased skin. Functionally, SDF-1 activates the ERK （extracellular-signal-regulated kinases） pathway and functions as a mitogen to stimulate epidermalkeratinocyte proliferation. Both overexpression of SDF-1 in dermal fibroblasts and treatment with rhSDF-1 to the skin equivalent cultures significantly increased the number of keratinocyte layers and epidermal thickness. Con- versely, the stimulative function of SDF-1 on keratinocyte proliferation was nearly completely eliminated by inter- fering with CXCR4, a specific receptor of SDF-1, or by knock-down of SDF-1 in fibroblasts. Our data reveal that extremely high levels of SDF-1 provide a crucial microenvironment for epidermal keratinocyte proliferation in both physiologic and pathologic skin conditions.

Protective effect of oat bran extracts on human dermal fibroblast injury induced by hydrogen peroxide

Oat contains different components that possess antioxidant properties; no study to date has addressed the antioxidant effect of the extract of oat bran on the cellular level. Therefore, the present study focuses on the investi- gation of the protective effect of oat bran extract by enzymatic hydrolysates on human dermal fibroblast injury induced by hydrogen peroxide （H2O2）. Kjeldahl determination, phenol-sulfuric acid method, and high-performance liquid chromatography （HPLC） analysis indicated that the enzymatic products of oat bran contain a protein amount of 71.93%, of which 97.43% are peptides with a molecular range from 438.56 to 1301.01 Da. Assays for 1,1-diphenyl-2-picrylhydrazyl （DPPH） radical scavenging activity indicate that oat peptide-dch extract has a direct and concentration-dependent antioxidant activity. 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide （MTT） colorimetric assay and the TdT-mediated digoxigenin-dUTP nick-end labeling （TUNEL） assay for apoptosis showed that administration of H2O2 in human dermal fibroblasts caused cell damage and apoptosis. Pre-incubation of human dermal fibroblasts with the Oatp for 24 h markedly inhibited human dermal fibroblast injury induced by H2O2, but ap- plication oat peptides with H2O2 at same time did not. Pre-treatment of human dermal fibroblasts with Oatp significantly reversed the H2O2-induced decrease of superoxide dismutase （SOD） and the inhibition of malondialdehyde （MDA）. The results demonstrate that oat peptides possess antioxidant activity and are effective against H2O2-induced human dermal fibroblast injury by the enhanced activity of SOD and decrease in MDA level. Our results suggest that oat bran will have the potential to be further explored as an antioxidant functional food in the prevention of aging-related skin injury.

Comparative study of the effects of gold and silver nanoparticles on the metabolism of human dermal fibroblasts

The purpose of this article was to explore the effects of gold nanoparticles(GNPs)and silver nano-particles(SNPs)with different cytotoxicities on human dermal fibroblasts(HDFs)at the metabolic level.First,~20 nm of GNPs and SNPs were prepared,and their effects on the proliferation of HDFs were evaluated.Then,a metabolomics technique was used to analyse the effects of GNPs and SNPs on the expression profiles of metabolites in HDFs after 4,8 and 24h of treatment.Furthermore,the key metabolites and key metabolic pathways involved in the interaction of GNPs and SNPs with HDFs were identified through expression pattern analysis and metabolic pathway analysis of differentially expressed metabolites and were finally verified by experiments.The results of the cytotoxicity experiments showed that there was no cytotoxicity after the treatment of GNPs for 72 h,while the cytotoxicity of the SNPs reached grade 1 after 72 h.By using metabolomics analysis,29,30 and 27 metabolites were shown to be differentially expressed in HDFs after GNP treatment,while SNPs induced the differential expression of 13,33 and 22 metabolites after 4,8 and 24h of treatment,respectively.Six and four candidate key metabolites in the GNP and SNP groups were identified by expression pattern analysis and metabolic pathway analysis,respec-tively.The key metabolic pathways in the GNP and SNP groups were identified as the glutathione metabolic pathway(the key metabolite of which was glutathione)and the citrate cycle pathway(the key metabolite of which was malic acid).Based on the experiments used to verify the key metabolites and key metabolic pathways,it was found that the increase in glutathione after GNP treatment might trigger an oxidative stress protection mechanism and thus avoid cytotoxicity.After exposure to SNPs,the citric acid content was increased,mainly through the citrate cycle path-way,thereby inhibiting the synthesis of malic acid to affect the formation of ATP and finally leading to cytotoxicity.

In vitro antioxidant and wound healing activity of Sargassum polycystum hydroethanolic extract in fibroblasts and keratinocytes

Objective:To investigate the in vitro antioxidant and wound healing properties of the hydroethanolic extract of Sargassum polycystum,and elucidate the mechanism of its wound healing activity.Methods:Human dermal fibroblast and HaCaT cells were used to evaluate the proliferation by sulforhodamine B and dsDNA assay after treatment with Sargassum polycystum extracts.Scratch wound healing and phalloidin-rhodamine staining were employed to observe migratory activity and filopodia formation,respectively.Western blot and real-time RT-PCR assays were performed to determine the protein and gene expressions related to wound healing activities.Results:The phytochemical analysis found a higher level of flavonoid than phenolic compound in Sargassum polycystum extracts.In human dermal fibroblast cells,Sargassum polycystum extracts at 50 and 100μg/mL significantly increased fibroblast proliferation and the gene expressions of hyaluronic acid synthase 1(HAS1),HAS2,HAS3,collagen type 1 alpha 1 chain(COL1A1),collagen type 3 alpha 1 chain(COL3A1),and elastin.The phosphorylation of Akt,ERK1/2,and p38 MAPK was also significantly upregulated after treatment with Sargassum polycystum extracts.Additionally,50 and 100μg/mL of the extracts prominently enhanced the proliferation,migration,and filopodia formation of HaCaT cells,as well as the protein levels of pFAK/FAK,pSrc/Src,pAkt/Akt,pERK1/2/ERK1/2,Rac1 and Cdc42.Conclusions:Sargassum polycystum extracts show promising wound healing activities in human dermal fibroblasts and keratinocytes.

Honey exposure stimulates wound repair of human dermal fibroblasts

Honey is widely used for treating burns, ulcers and wounds, but the mechanisms of action are poorly known and the product is mainlyused as an antimicrobial. We have examined here the wound healing properties of honey on human fi broblasts, using an in vitroscratch wound healing model. Three kinds of widely used monofl oral honeys were used, viz. acacia (Robinia pseudacacia), buckwheat(Fagopyrum sp.), and manuka (Leptospermum scoparium). Data displayed an increased wound healing activity in fi broblasts, butwith diff erent effi ciency and mechanisms of action among honeys. The eff ects of acacia and buckwheat emerged in both scratchwound and chemotaxis assays, while the eff ect of manuka was signifi cant but lower. The use of inhibitors indicated on the wholean essential role of cytosolic calcium, an important role of ERK and p38, and a secondary role of PI3K. Acacia and buckwheat,but not manuka, induced signifi cant increases in the release of interleukin-4 (IL-4), IL-6, and IL-8, indicating a correlation betweeninterleukin upregulation and wound closure effi ciency. This is consistent with our previous fi ndings suggesting a higher ability ofacacia and buckwheat to activate keratinocyte reepithelialization, with respect to manuka honey. In conclusion, our data indicatethat acacia and buckwheat honeys are particularly effi cient in facilitating fi broblast wound closure activities, suggesting newtherapeutic possibilities for this natural product.

UV-induced senescence of human dermal fibroblasts restrained by low-stiffness matrix by inhibiting NF-κB activation

As a hallmark of skin aging,senescent human dermal fibroblasts(HDFs)are known to lose the ability to divide.However,they can still interact with their cellular environment and the surrounding matrix.As the skin ages,the progressive slowing down of HDFs function decreases the skin’s structural integrity,which is more serious than if there is the dermal collagen matrix eroded.This leads to matters of the unbalanced barrier under the skin,skin fragility,inadequate wound healing,as well as other cosmetic issues.It is also well documented that skin aging comes with significant stiffness increases.Therefore,understanding the interactions between HDFs and the surrounding microenvironments during senescence may provide insights into skin aging.Here we aim to inves-tigate matrix stiffness’effect on HDF senescence and elucidate possible mechanisms that make HDFs senescent.In our experiments,HDFs were cultivated on Polydimethylsiloxane(PDMS)with various stiffnesses and exposed to UV light to trigger senescence.Results show that HDFs are significantly affected by senescence when cultured on a matrix with stiffness.However,the cells are not significantly affected when cultured on a low stiffness matrix.The following characterization revealed cells cultured on stiffsubstrates under UV exposure had stimu-lated the nucleus factor kappa-B(NF-κB)activation.In contrast,cells on a matrix of softness only displayed low activation of NF-κB.NF-κB activity suppression with ammonium pyrrolidine dithiocarbamate(PDTC)decreases UV-induced HDFs senescence on stiffsubstrates.Taken together,we demonstrated that soft matrix defends HDFs against ultraviolet-induced senescence by inhibiting the activation of NF-κB.

Human dermal reticular fibroblasts at confluence display a signature micro pattern <i>in vitro</i>

This paper sets out to demonstrate that scraping of the flat dorsal surface of human dermis with a scalpel blade and cell plating without centrifugation can lead to the recognition and identification of the individual packing micro pattern of dermal reticular fibroblasts at confluence. The characteristic alignment of papillary and reticular fibroblasts at right angles to each other led to the positive identification of reticular fibroblasts. A non-enzymatic means of sub-culturing (passaging), which yields fully functional, healthy cells with normal, phenotypic morphology is also described. Implications for published subcutaneous wound healing studies are discussed as well as the confluent reticular fibroblast configuration, interpreted as ananatomic site identity code,which may be the address of a specific fibroblast gene pattern expression.

<i>In Vitro</i>Analysis of VEGF and HGF Production by Fibroblast in Cultured Dermal Substitute Combined with EGF-Incorporating Top Dressing

This study aimed to investigate the potential of cultured dermal substitute (CDS) to release angiogenic growth factors when laminated with a membrane containing epidermal growth factor (EGF) as a top dressing. Membranes were prepared by air-drying a solution of hyaluronic acid (HA) and collagen (Col) with or without EGF. Membranes were designed to contain EGF at concentrations of 0, 0.1, 0.2 or 0.5 μg/cm2. CDS was prepared by incorporating fibroblasts into a collagen gel combined with a cross-linked HA spongy matrix, followed by culturing for 5 days. CDS was designed to contain fibroblasts at a density of 2 × 105 (Group I) or 4 × 105 cells/cm2> (Group II). CDS was elevated at the interface between air and culture medium, on the top of which each membrane was placed. This culture system was employed as a wound surface model. Metabolic activity of the fibroblasts in the CDS cultured for 7 days on a wound surface model was measured by MTT assay. The amounts of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) after 7 days of cultivation were measured by using ELISA. Membranes containing EGF ranging from 0.1 to 0.5 μg/cm2> facilitated production of both VEGF and HGF, as compared with control membranes without EGF. However, a membrane containing EGF at a concentration of 0.5 μg/cm2> failed to facilitate fibroblast cytokine production in Group I. These results demonstrated that the EGF-incorporating membrane was able to stimulate fibroblasts in the CDS to synthesize an increased amount of VEGF and HGF in a dose-dependent manner.

不同来源成纤维细胞对角质形成细胞再上皮化影响

目的比较人牙龈及皮肤来源成纤维细胞条件培养基对永生化角质形成细胞再上皮化作用的差异,分析成纤维细胞促进再上皮化作用的关键因子,探究牙龈来源成纤维细胞在创伤修复中的可能应用。方法分别制备人牙龈成纤维细胞(hGFs)和人真皮成纤维细胞(hDFs)条件培养基(CM),添加到人永生化角质形成细胞(Human keratinocyte cells,HaCaT)中,采用CCK-8、EdU染色检测HaCaT细胞增殖能力;细胞划痕实验、Transwell细胞迁移实验检测细胞迁移能力;通过ABplex多因子检测及酶联免疫吸附法(ELISA)对hGFs、hDFs分泌的多种与创伤修复相关的生长因子、细胞因子和趋化因子进行定量分析。结果CCK-8、EdU染色、细胞划痕实验、Transwell细胞迁移实验结果表明:2种条件培养基均能显著提高HaCaT细胞的增殖及迁移能力(P<0.05),hGFs-CM对HaCaT细胞增殖和迁移能力的促进作用显著强于hDFs-CM(P<0.01);hGFs和hDFs分泌的因子有较大的差异,其中hGFs分泌的生长因子如肝细胞生长因子(HGF)、血管内皮生长因子(VEGF)明显多于hDFs。结论人牙龈来源成纤维细胞较人皮肤来源成纤维细胞的条件培养基对角质形成细胞的增殖和迁移能力的有更强的促进作用,可能与其旁分泌产生的因子不同有关。

年轻化间充质干细胞条件培养基促进小鼠创面愈合的研究

背景间充质干细胞(mesenchymal stem cell,MSC)条件培养基无细胞疗法是创伤修复领域的一种新型治疗方案,但其治疗效果受间充质干细胞状态的影响。目的探究年轻化MSC培养基对小鼠创面愈合的作用。方法建立间充质干细胞复制性衰老(传代至15~20代)模型,检测其衰老指征;使用小分子化合物[丙戊酸(valproic acid,VPA)和RepSox]逆转衰老间充质干细胞并验证逆衰老效果。分别收集衰老MSC培养基与年轻化MSC培养基。用收集到的两种条件培养基培养人成纤维细胞,使用CCK-8实验、Ki-67免疫荧光、划痕实验和Transwell实验检测这两种条件培养基对成纤维细胞增殖、迁移和侵袭能力的影响。分别用衰老MSC培养基、年轻化MSC培养基和0.9%氯化钠注射液治疗C57BL/6小鼠背部皮肤全层缺损模型,评估3种治疗方法对创面愈合的影响。结果与衰老的间充质干细胞相比,经小分子处理后衰老间充质干细胞SA-β-gal阳性率降低(P<0.01),且免疫荧光检测显示其衰老基因P21、P16表达降低(P<0.01)。经年轻化MSC培养基处理的成纤维细胞增殖、迁移和侵袭能力增强(P<0.01)。创伤模型显示,与对照组相比,年轻化MSC培养基治疗组小鼠创面愈合速度加快(P<0.01);年轻化MSC培养基治疗组小鼠创面HE染色显示表皮细胞再上皮化加快(P<0.01)、Masson染色显示创面胶原沉积量增加(P<0.01)、CD31免疫荧光染色显示创面血管量增加(P<0.01)。结论年轻化MSC培养基对小鼠创面修复或有促进作用。

口服生物活性胶原蛋白肽的筛选与体外功效评价

本文研究9个胶原蛋白肽的溶解度、羟脯氨酸含量、肽相对分子量和促进人真皮成纤维细胞生长功效四个方面的特性,为开发人真皮成纤维细胞生长的生物活性胶原蛋白肽功能性饮品提供参考。

Hyaluronate Increases Polynucleotides Effect on Human Cultured Fibroblasts

The HA is present in almost all vertebrates and plays a critical role in tissue development and cell proliferation, it has been demonstrated to promote wound healing and involved in angiogenesis and inflammation. Also polynucleotydes (PN) have proved to promote the “in vitro” growth and activity of human fibroblasts and osteoblasts, to increase reparation on UVB damaged dermal fibroblasts and seems to promote proliferation of human pre-adipocytes. Several in vivo studies have demonstrated the PN effect also in vivo, inducing an increase of angiogenesis and healing process. In this paper we have evaluated the effect of a mixture of Polynucleotides (PN) and entire Hyaluronic Acid (HA) on cultured human fibroblasts by analyzing cell growth. Different mixture have been tested and it has been demonstrated that the presence of HA even at low concentration (1 mg/ml) determine an increase of PN activity up to 20%. Furthermore, the addition of HA 1 mg/ml to PN 100 μg/ml induces a cell growth rate comparable to that exerted by PN concentration of 12 μg/ml.

Scientific Viewpoints with Emphasis on Dermal Cellular Regeneration in Wound Sites

The human dermis presents an ongoing problem for regenerative medicine. Current medical management uses various acellular dermal matrices on wound sites. The challenge for scientists is to examine, then to question accepted conventional wisdom and to present new concepts. In this paper, Autologous Cell Therapy will be described by using cell culture of autologous dermal fibroblasts and their extracellular matrix as a foundation for rebuilding the dermis in conditioned wound beds. This proposal seems to create a conflict with the medical approach to keeping a wound bed “moist”.