title: Engineering of implantable cartilaginous structures from bone marrow-derived mesenchymal stem cells.
abstract: Fabrication of implantable cartilaginous structures that could be secured in the joint defect could provide an alternative therapeutic approach to prosthetic joint replacement. Herein we explored the possibility of using biodegradable hydrogels in combination with a polyglycolic acid (PGA) scaffold to provide an environment propitious to mesenchymal stem cells (MSCs) chondrogenic differentiation. We examined the influence of type I collagen gel and alginate combined with PGA meshes on the extracellular matrix composition of tissue-engineered transplants. MSCs were isolated from young rabbits, expanded in monolayers, suspended in each hydrogel, and loaded on PGA scaffolds. All constructs (n=48) were cultured in serum-free medium containing transforming growth factor beta-1, under dynamic conditions in specially designed bioreactors for 3-6 weeks. All cell-polymer constructs had a white, shiny aspect, and retained their initial size and shape over the culture period. Their thickness increased substantially over time, and no shrinkage was observed. All specimens developed a hyalin-like extracellular matrix containing glycosaminoglycans (GAGs) and type II collagen, but significant differences were observed among the three different groups. In PGA/MSCs and collagen-PGA/MSCs constructs, the cell growth phase and the chondrogenic differentiation phase of MSCs occurred during the first 3 weeks. In alginate-PGA/MSCs constructs, cells remained round in the hydrogel and cartilage extracellular matrix deposition was delayed. However, at 6 weeks, alginate-PGA/MSCs constructs exhibited higher contents of GAGs and lower contents of type I collagen. These results suggest that the implied time for the transplantation of in vitro engineered constructs depends, among other factors, on the nature of the scaffold envisioned. In this study, we demonstrated that the use of a composite hydrogel-PGA scaffold supported the in vitro growth of implantable cartilaginous structures cultured in a bioreactor system.
authors.affiliation: Laboratory for Tissue Engineering and Organ Fabrication, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA.
authors.forename: D
authors.initials: D
authors.lastname: Hannouche
authors.forename: H
authors.initials: H
authors.lastname: Terai
authors.forename: J R
authors.initials: JR
authors.lastname: Fuchs
authors.forename: S
authors.initials: S
authors.lastname: Terada
authors.forename: S
authors.initials: S
authors.lastname: Zand
authors.forename: B A
authors.initials: BA
authors.lastname: Nasseri
authors.forename: H
authors.initials: H
authors.lastname: Petite
authors.forename: L
authors.initials: L
authors.lastname: Sedel
authors.forename: J P
authors.initials: JP
authors.lastname: Vacanti
mesh_terms: D000464:Alginates / Q000648:ultrastructure; D000818:Animals; D001672:Biocompatible Materials / Q000138:chemical synthesis; D019149:Bioreactors; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology / Q000648:ultrastructure; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques; D024042:Collagen Type I / Q000138:chemical synthesis / Q000648:ultrastructure; D024043:Collagen Type II / Q000138:chemical synthesis / Q000648:ultrastructure; D020723:Glucuronic Acid / Q000502:physiology; D006603:Hexuronic Acids; D051457:Hyaline Cartilage / Q000502:physiology / Q000637:transplantation* / Q000648:ultrastructure; D020100:Hydrogels; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D011100:Polyglycolic Acid; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D000464:Alginates; D001672:Biocompatible Materials; D024042:Collagen Type I; D024043:Collagen Type II; D006603:Hexuronic Acids; D020100:Hydrogels; D011100:Polyglycolic Acid; D020723:Glucuronic Acid