title: The use of gene activated matrix to mediate effective SMAD2 gene silencing against hypertrophic scar.
abstract: Hypertrophic scar (HS) originates from the over-expression of transforming growth factor β (TGF-β) and downstream SMAD2. With attempts to rectify HS by RNA interference (RNAi) against SMAD2, we report the design of plasmid DNA encoding SMAD2 siRNA (pSUPER-SMAD2), and identify the optimal siRNA sequence toward maximal RNAi efficiency. To realize effective and sustained RNAi, we developed gene activated matrix (GAM) based on porous atelocollagen scaffold and embedded trimethyl chitosan-cysteine (TMCC)/pSUPER-SMAD2 polyplexes for promoting cell growth and gene transfection. The GAM exhibited porosity higher than 80%, pore size of 200-250 μm, desired mechanical strength, and sustained pSUPER-SMAD2 release profiles. Normal skin fibroblasts (NSFs) and hypertrophic scar fibroblasts (HSFs) were allowed to infiltrate and proliferate in GAM; at the meantime they were transfected with TMCC/pSUPER-SMAD2 polyplexes to display remarkably reduced SMAD2 levels that lasted for up to 10 days, consequently inhibiting the over-production of type I and type III collagen. We further unraveled the notably higher transfection levels of GAM in three-dimensional (3D) than in 2D environment, which was attributed to the improved cell-matrix interactions that promote cell proliferation and polyplex internalization. This highly safe and effective GAM may serve as a promising candidate towards HS treatment.
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Pharmaceutical Sciences, School of Life Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Lichen
authors.initials: L
authors.lastname: Yin
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Pharmaceutical Sciences, School of Life Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Center of Burns and Traumatic Surgery, Changhai Hospital, Second Military Medical University, Shanghai 200433, China.
authors.forename: Shizhao
authors.initials: S
authors.lastname: Ji
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Pharmaceutical Sciences, School of Life Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Chunbai
authors.initials: C
authors.lastname: He
authors.affiliation: Center of Burns and Traumatic Surgery, Changhai Hospital, Second Military Medical University, Shanghai 200433, China. Electronic address: southeast000@163.com.
authors.forename: Guangyi
authors.initials: G
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Pharmaceutical Sciences, School of Life Sciences, Fudan University, Shanghai 200433, China. Electronic address: tangcui@fudan.edu.cn.
authors.forename: Cui
authors.initials: C
authors.lastname: Tang
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Pharmaceutical Sciences, School of Life Sciences, Fudan University, Shanghai 200433, China. Electronic address: shaohuagu@fudan.edu.cn.
authors.forename: Shaohua
authors.initials: S
authors.lastname: Gu
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Pharmaceutical Sciences, School of Life Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Chunhua
authors.initials: C
authors.lastname: Yin
mesh_terms: D001483:Base Sequence; D049109:Cell Proliferation; D017439:Cicatrix, Hypertrophic / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D003094:Collagen / Q000378:metabolism; D004247:DNA / Q000235:genetics; D015536:Down-Regulation; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction; D012867:Skin / Q000166:cytology / Q000473:pathology; D051899:Smad2 Protein / Q000235:genetics* / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; C497782:SMAD2 protein, human; D051899:Smad2 Protein; D016212:Transforming Growth Factor beta; C059821:atelocollagen; D003094:Collagen; D004247:DNA
keywords: Atelocollagen; Gene activated matrix (GAM); Hypertrophic scar; RNA interference (RNAi); Thiolated trimethyl chitosan