Skin care efficacy study of recombinant humanized collagen based on in vitro level

Studying the skin care efficacy of recombinant humanized collagen based on in vitro level.The stability of the recombinant humanized collagen was first analyzed by treating at different temperatures,then its skincare efficacy based on in vitro level was evaluated by detecting the inhibition rate of elastase,the inhibition rate of collagenase,the protein content of type I collagen in human fibroblasts,the inhibition of reactive oxygen species(ROS)with human keratinocytes,and the effects of the recombinant humanized collagen on the expression of hyaluronic acid(HA),filaggrin(FLG)and transglutaminase 1(TGM1)in keratinocytes.The results showed that recombinant humanized collagen was able to maintain stability at temperatures below 70℃.With regard to its skincare efficacy,recombinant humanized collagen could inhibit elastase and collagenase activities and promote the increase of type I collagen content in human fibroblasts.It also showed good inhibition of ROS in keratinocytes in vitro and could increase the expression of HA,FLG,and TGM1 in keratinocytes.In short,the recombinant humanized collagen exhibited a favourable skin care effect in vitro level.This study proved that it has potential firming,anti-wrinkle,moisturizing,and repairing efficacy,and is a valuable cosmetic raw material.

Recombinant humanized collagen remodels endometrial immune microenvironment of chronic endometritis through macrophage immunomodulation

Recently,evidence has suggested that chronic endometritis(CE)is a crucial factor associated with infertility and failure of assisted reproductive techniques,prompting concern in the reproductive field.Studies have shown that persistent infiltered immune cells stimulation result in the disturbance of endometrial immune microenvironment could lead to the infertility of CE patients finally.Conventional treatments are limited because they lack immune regulation,so it is urgent to develop a novel approach to treat CE and promote embryo implantation in patients with CE.Herein,we prepared recombinant humanized type III collagen(rhCol III)with high cell adhesion activity to regulate macrophages and repair the endometrium.In this study,M1 macrophages and M1 macrophages cultured medium and lipopolysaccharide(LPS)co-stimulated inflammatory endometrium stromal cells(ESCs)were established in vitro to mimic CE condition.rhCol III promoted M1 macrophages toward M2 phenotype,improved cell migration,viability and collagen components of inflammatory ESCs.Also,the inflammatory response of inflammatory ESCs was downregulated after rhCol III treatment.Subsequently,LPS was used for CE rat model and a 28-day observation was performed;inflammatory cells’infiltration,endometrium repair,extracellular matrix(ECM)remodeling and pregnancy outcomes were promoted after rhCol III endometrial infusion.In conclusion,rhCol III promoted(i)macrophage polarization toward M2 macrophages,(ii)pro-inflammatory cytokine production and anti-inflammatory cytokine reduction,(iii)ECM remodeling and(iv)fertility restoration.Meanwhile,rhCol III enhanced cell biological functions by interacting with discoidin domain receptors,regulated cell metabolism and reduced the inflammatory response through the inhibition of the NF-κB/YAP signaling pathway.Overall,the results illustrated the potential therapeutic prospects of rhCol III for CE treatment.

The influence of tag sequence on recombinant humanized collagen(rhCol)and the evaluation of rhCol on Schwann cell behaviors

Recombinant humanized collagen(rhCol)was an extracellular matrix(ECM)-inspired biomimetic biomaterial prepared by biosynthesis technology,which was considered non-allergenic and could possibly activate tissue regeneration.The influence of tag sequence on both structures and performances of rhCol type III(rhCol III)was investigated,and the effect of rhCol III on cell behaviors was evaluated and discussed using Schwann cells(SCs)as in vitro model that was critical in the repair process after peripheral nerve injury.The results demonstrated that the introduction of tag sequence would influence both advanced structures and properties of rhCol III,while rhCol III regulated SCs adhesion,spreading,migration and proliferation.Also,both nerve growth factor and brain-derived neurotrophic factor increased when exposed to rhCol III.As the downstream proteins of integrin-mediated cell adhesions,phosphorylation of focal adhesion kinase and expression of vinculin was up-regulated along with the promotion of SCs adhesion and migration.The current findings contributed to a better knowledge of the interactions between rhCol III and SCs,and further offered a theoretical and experimental foundation for the development of rhCol III-based medical devices and clinical management of peripheral nerve injury.

Ceruloplasmin or Fibronectin Synergism with Quartz Dust on Stimulating Collagen Gene Transcription in Human 2BS Fibroblast

Human α1(Ⅰ), α2(Ⅰ) and α1(Ⅲ) cDNA probes and RNA dot hybridization were employed to quantitate collagen mRNA changes after adding silica dust into the media of human 2BS fibroblasts. At all dosages used (100, 200, 500 and 1000μg), the α1(Ⅰ), α2(Ⅰ)and α1(Ⅲ) mRNA levels increased one day after dusting. At the same dosage of silica (100μg), α1(Ⅲ) mRNA increased earlier than type Ⅰ collagen mRNA did. The type Ⅰ and type Ⅲ collagen mRNA contents in the experimental groups were higher than those in control on days 3, 5, 7 and 9. The effect of ceruloplasmin (Cp) and fibronectin (Fn) on collagen mRNA synthesis was also studied, after adding silica dust, Cp or Fn into the media of human 2BS fibroblast. The results showed that Cp and Fn have stimulating effect on collagen mRNA production. When both Cp and silica dust were added into cell culture media, the collagen mRNA level was increased more than those of adding either Cp or silica dust alone. Similar situations were found for Fn. Cp (or Fn) synergism with silica dust on stimulating transcription of human collagen gene was suggested

Experimental study on effect of endothelin in the proliferation and collagen synthesis of human scar-derived fibroblasts

Objective To investigate the role of endothelin(ET) in the proliferation and collagen synthesis of human scar-derived fibroblasts and the moduktion of its antagonists such as nitric oxide(NO), tetrandrine ( Tet). Methods With the cultured fibroblasts from the scarring tissue, the cell pdiferation was determined by[3H]-TdR incorporation, while the collagen synthesis was evaluated by[3H]-proline incorporation. Results The ET-1 was significantly increasing the proliferation and collagen synthesis of human scar-derived fibroblasts. The values of [3H]-TdR absorption in the 2.5 ng/ml,25 ng/ml and 100 ng/ml of ET-1 groups were 1.8 times,4 times and 4.9 times more than in the control group, respectively(P【0. 01),while the values of the [3H]-proline incorporation were 1.1 times,3.1 times and 3.8 times respectively(P【0.01). The fibroblasts, treated with 50 μg/ml of S-nitroso-N-acetyl penicillamine(SNAP), were no detectable effect on the basal level of DNA synthesis,but produced decreasing effect on the

A strategy for mechanically integrating robust hydrogeltissue hybrid to promote the anti-calcification and endothelialization of bioprosthetic heart valve

Bioprosthetic heart valve(BHV)replacement has been the predo-minant treatment for severe heart valve diseases over decades.Most clinically available BHVs are crosslinked by glutaraldehyde(GLUT),while the high toxicity of residual GLUT could initiate calcification,severe thrombosis,and delayed endothelializa-tion.Here,we construed a mechanically integrating robust hydrogel-tissue hybrid to improve the performance of BHVs.In particular,recombinant humanized coilagen type Ⅲ(rhCOLⅢ),which was precisely customized with anti-coagulant and pro-endothelialization bioactivity,was first incorporated into the polyvinyl alcohol(PVA)-based hydrogel via hydrogen bond interactions.Then,tannic acid was introduced to enhance the mechanicalperfo of PVA-based hvdrogel and interfacial bonding between the hydrogel layer and bio-derived tissue due to the strong affinity for a wide range of substrates.In vitro and in vivo experimental results confirmed that the GLUT-crosslinked BHVs modified by the robust PVA-based hydrogel embedded rhCOLII and TA possessed long-term anti-coagulant,accelerated endothelialization,mild inflammatory response and anti-calcification properties.Therefore,our mechanically integrating robust hydrogel-tissue hybrid strategy showed the potential to enhance the service function and prolong the service life of the BHVs after implantation.

An extracellular matrix-mimetic coating with dual bionics for cardiovascular stents

Anti-inflammation and anti-coagulation are the primary requirements for cardiovascular stents and also the widely accepted trajectory for multi-functional modification.In this work,we proposed an extracellular matrix(ECM)-mimetic coating for cardiovascular stents with the amplified functionalization of recombinant humanized collagen type III(rhCOL III),where the biomimetics were driven by structure mimicry and component/function mimicry.Briefly,the structure-mimic was constructed by the formation of a nanofiber(NF)structure via the polymerization of polysiloxane with a further introduction of amine groups as the nanofibrous layer.The fiber network could function as a three-dimensional reservoir to support the amplified immobilization of rhCoL III.The rhCOL III was tailored for anti-coagulant,anti-inflammatory and endothelialization promotion properties,which endows the ECM-mimetic coating with desired surface functionalities.Stent implantation in the abdominal aorta of rabbits was conducted to validate the in vivo re-endothelialization of the ECM-mimetic coating.The mild inflammatory responses,anti-thrombotic property,promotion of endothelialization and suppression of excessive neointimal hyperplasia confirmed that the ECM-mimetic coating provided a promising approach for the modification of vascular implants.