Halomonas levan for cosmeceutical applications

ÖZETCilt, vücudu su kaybından ve bakteri penetrasyonu gibi dış etkenlerden koruyan endıştaki tabakadır. Hyaluronik asit, kolajen, filaggrin veya involukrin proteinlerindemeydana gelen azalma su kaybı, kırışıklık oluşumu, cilde bakteri girişi, atopik dermatit,sedef hatalığı gibi pek çok rahatsızlığa sebep olmaktadır. Biyolojik olarak aktif doğalbileşenler içeren kozmetiklere artan ilgi biyoaktif, biyouyumlu ve biyobozunurpolisakkaritlerin kozmetik endüstrisince cilt bakım ürünlerinde kullanılmak üzere tercihedilmesini arttırmıştır. Hyaluronik asit, kolajen, selüloz ve poli-gama-glutamik asit aktifdoğal bileşenler olarak sıklıkla kullanılmaktadır. Doğada en yaygın bulunan fruktan türüolan β (2→6)- bağlı levan polisakkariti, düşük toksisite, biyouyumluluk, film oluşturmave yüksek suda çözünürlük gibi özellikleri sebebiyle kimya, ecza, kozmetik ve gıdaendüstrilerince sıklıkla tercih edilmektedir. İlk levan üreten halofilik bakteri Halomonassmyrnensis AAD6T’nin ürettiği levanın anti-kanser, heparin mimetik, hücreproliferasyonunu destekleyen özellikleri daha önce bildirilmiştir. Bu doktora tezikapsamında, Halomonas levan, kimyasal değişikliğe uğratılmış türevleri, dallanmışBacillus subtilis levan, Chromohalobacter canadensis 28 ekzopolimeri (Cc EP) vesaflaştırılmış fraksiyonu olan poli-gama-glutamik asit (γ-PGA) polimerinin insankeratinosit (HaCaT) ve fibroblast (PCS-201-012) hücreleri ile hücre canlılığı, kolajen,hyaluronik asit, involukrin, filaggrin gen anlatımına etkisi, hücre siklus analizi ve yarakapanması üzerine etkilerinin araştırılması hedeflenmiştir. Testler sonucunda HLpolimerinin, kimyasal türevleri, Cc EP ve PGA, hyaluronik asitin, kolajen, filaggrin veinvolükrin gen anlatımını tetikleyen ve yara kapanması ile hücre proliferasyonunudestekleyen özellikleri sayesinde kozmesötik bileşen olarak kullanılabilmepotansiyeline sahip olduğu kararlaştırılmıştır.--------------------ABSTRACTSkin protect inside from external potential damages like water loss or bacterialpenetration. Changes in this tissues like decrease in collagen, hyaluronic acid or elastinproduction or impaired barrier function due to filaggrin or involucrin decrease result inwrinkle formation, water loss, bacterial penetration, diseases like atopic dermatitis,ichthyosis, psoriasis, acne and aged skin look with fine or deep wrinkles. Demand onnatural products that contain biological ingredients with mimicking naturally occurringgrowth factors and cytokines for several biological metabolisms made naturalpolysaccharide popular in cosmetic industry. Hyaluronic acid, collagen, cellulose andpoly--glutamic acid (γ-PGA)are used as active ingredients. Levan is a β (2→6)- linked,one of the most common fructan polymer in nature. Water soluble, film former, stronglyadhesive, non-toxic, biocompatible and bioactive levan polysaccharide has beenreported a novel polymer for various application fields including cosmetics, food,medicine, pharmacy and chemical industries. Halophilic bacterium Halomonassmyrnensis AAD6T is defined as the first levan producer extremophile with variousbioactivities reported before. Here in this thesis investigation, Halomonas smyrnensislevan (HL), its chemically derived forms, branched levan from Bacillus subtilis,Chromohalobacter canadensis 28 exopolymer (Cc EP) and purified fraction of polygamma glutamic acid (γ-PGA) are tested with human keratinocytes (HaCaT) andfibroblast cells (PS-201-012) for cellular proliferation, collagen, hyaluronic acid,involucrin and filaggrin gene expression, cell cycle analysis, wound closure effect on invitro scratch assay and concluded as an active ingredient for cosmeceuticals with theircell proliferative, collagen, hyaluronic acid, involucrin and filaggrin expressionpromoting and wound closing properties

Improved Exopolymer Production by Chromohalobacter canadensis Cultures for Its Potential Cosmeceutical Applications

Several exopolymers with different chemical composition and correspondingly variety in their physico-chemical properties from halophilic microorganisms have still been described, however, with a low production yield. Chromohalobacter canadensis 28 isolated from Pomorie saltern synthesized an unusual exopolymer (EP) containing 72% γ-polyglutamic acid (PGA), an essential cosmeceutical additive. Current work suggests a novel approach for effective EP synthesis by C. canadensis 28 using continuous cultures. Highest production was observed at low dilution rates reaching a level of 2.1 mg/mL at D = 0.035, similar to those in batch cultures (2.34 mg/mL), however avoiding all disadvantages of discontinuous fermentation processes. At steady state, the total quantities of the synthesized EP after 48 h cultivation for the given equipment volume in D = 0.035 h−1 and D = 0.075 h−1 were 8.67 and 12 g, correspondingly, while it was 2.9 g for batch culture. Process parameters did not change after a ten-day run at D = 0.35 h−1. A degree of purity of EP fraction received from continuous cultures was significantly increased up to 93–96%. A lack of cytotoxicity and high cell viability were observed for human dermal fibroblast cells after 24 h incubation with crude EP from C. canadensis 28 and purified PGA fraction that could suggest its high potential for cosmetic applications

Bioinspired scaffold induced regeneration of neural tissue

WOS: 000467668800012In the last decade, nerve tissue engineering has attracted much attention due to the incapability of self-regeneration. Nerve tissue regeneration is mainly based on scaffold induced nanofibrous structures using both bio and synthetic polymers. The produced nanofibrous scaffolds have to be similar to the natural extracellular matrix and should provide an appropriate environment for cells to attach onto. Nanofibrous scaffolds can support or regenerate cells of tissue. Electrospinning is an ideal method for producing the nanofibrous scaffolds. In this study, Bacterial cellulose (BC)/Poly (epsilon-caprolactone) (PCL) blend nanofibrous scaffolds were successfully prepared by electrospinning for nerve tissue induced repair. The produced nanofibrous scaffolds contain well defined interconnected nanofiber networks with hollow micro/nanobeads. Firstly, in-vitro biocompatibilities of nanofibrous scaffolds were tested with L2929 murine fibroblasts and improved cell adhesion and proliferation was observed with polymer blends compared with PCL only. The primary cell culture was performed with dorsal root ganglia (DRG) cells on nanofibrous samples and the samples were found suitable for enhancing neural growth and neurite outgrowth. Based on these results, the BC/PCL (50:50 wt%) nanofibrous scaffolds exhibited nerve-like branching and are excellent candidate for potential biomimetic applications in nerve tissue engineering regeneration