title: Immobilized lentivirus vector on chondroitin sulfate-hyaluronate acid-silk fibroin hybrid scaffold for tissue-engineered ligament-bone junction.
abstract: The lack of a fibrocartilage layer between graft and bone remains the leading cause of graft failure after anterior cruciate ligament (ACL) reconstruction. The objective of this study was to develop a gene-modified silk cable-reinforced chondroitin sulfate-hyaluronate acid-silk fibroin (CHS) hybrid scaffold for reconstructing the fibrocartilage layer. The scaffold was fabricated by lyophilizing the CHS mixture with braided silk cables. The scanning electronic microscopy (SEM) showed that microporous CHS sponges were formed around silk cables. Each end of scaffold was modified with lentiviral-mediated transforming growth factor- β 3 (TGF- β 3) gene. The cells on scaffold were transfected by bonded lentivirus. In vitro culture demonstrated that mesenchymal stem cells (MSCs) on scaffolds proliferated vigorously and produced abundant collagen. The transcription levels of cartilage-specific genes also increased with culture time. After 2 weeks, the MSCs were distributed uniformly throughout scaffold. Deposited collagen was also found to increase. The chondral differentiation of MSCs was verified by expressions of collagen II and TGF- β 3 genes in mRNA and protein level. Histology also confirmed the production of cartilage extracellular matrix (ECM) components. The results demonstrated that gene-modified silk cable-reinforced CHS scaffold was capable of supporting cell proliferation and differentiation to reconstruct the cartilage layer of interface.
authors.affiliation: Institute of Orthopedic Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Liguo
authors.initials: L
authors.lastname: Sun
authors.affiliation: Institute of Orthopedic Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Hongguo
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Clinical Laboratory, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Qu
authors.affiliation: College of Science, Air Force Engineering University, Xi'an 710051, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Zhu
authors.affiliation: Institute of Orthopedic Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Xiangli
authors.initials: X
authors.lastname: Fan
authors.affiliation: Institute of Orthopedic Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Yingsen
authors.initials: Y
authors.lastname: Xue
authors.affiliation: Institute of Orthopedic Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Zhenghong
authors.initials: Z
authors.lastname: Xie
authors.affiliation: Institute of Orthopedic Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Hongbin
authors.initials: H
authors.lastname: Fan
mesh_terms: D000327:Adsorption; D000818:Animals; D001672:Biocompatible Materials / Q000138:chemical synthesis; D001842:Bone and Bones / Q000166:cytology / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D002809:Chondroitin Sulfates / Q000737:chemistry; D005349:Fibroins / Q000737:chemistry; D005822:Genetic Vectors / Q000235:genetics; D006820:Hyaluronic Acid / Q000737:chemistry; D016086:Lentivirus / Q000235:genetics*; D008022:Ligaments / Q000166:cytology / Q000502:physiology; D008422:Materials Testing; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology*; D011817:Rabbits; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods; D054457:Tissue Scaffolds; D053782:Transforming Growth Factor beta3 / Q000235:genetics*
chemical_list: D001672:Biocompatible Materials; D053782:Transforming Growth Factor beta3; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates; D005349:Fibroins