In vivo evaluation of MMP sensitive high-molecular weight HA-based hydrogels for bone tissue engineering.
Hyaluronic acid (170 kDa)-based hydrogel was synthesized using acrylated hyaluronic acid (HA) and matrix metalloproteinase (MMP) sensitive HA-based hydrogels were then prepared by conjugation with two different peptides: cell adhesion peptides containing integrin-binding domains (Arg-Gly-Asp: RGD) and a cross-linker with MMP degradable peptides to mimic the remodeling characteristics of natural extracellular matrices by cell-derived MMPs. Mechanical properties of these hydrogels were evaluated with different weight percentages (2.5 and 3.5 wt %) by measuring elastic modulus, viscous modulus, and swelling ratio. Human mesenchymal stem cells (hMSCs) were then cultured in MMP-sensitive or insensitive HA-based hydrogels and/or immobilized cell adhesive RGD peptides in vitro. Actin staining and image analysis proved that cells cultured in the MMP-sensitive hydrogel with RGD peptides showed extensive cell spreading and sprouting. Gene expression analysis showed that bone specific genes such as alkaline phosphatase, osteocalcin, and osteopontin increased in MMP-sensitive hydrogels as biomolecules such as BMPs and cells were added in the gels. For in vivo calvarial defect regeneration, five different samples (MMP insensitive hydrogel, MMP sensitive hydrogel, MMP sensitive hydrogel with BMP-2, MMP sensitive hydrogel with hMSC, and MMP sensitive hydrogel with BMP-2 and hMSC) were prepared. After 4 weeks of implantation, the Masson-Trichrome staining and micro computed tomography scan results demonstrated that the MMP sensitive hydrogels with BMP-2 and hMSCs have the highest mature bone formation. The MMP sensitive HA-based hydrogel could become useful scaffolds in bone tissue engineering with improvements on tissue remodeling rates and regeneration activity.
Kim
Jungju
J
Biomedical Engineering, Brain Korea 21, 2nd Project for Biomedical Science, Korea University Medical College, Korea University, Seoul 136-705, Korea.
Kim
In Sook
IS
Cho
Tae Hyung
TH
Kim
Ho Chul
HC
Yoon
So Jeong
SJ
Choi
Jaesoon
J
Park
Yongdoo
Y
Sun
Kyung
K
Hwang
Soon Jung
SJ
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry / Q000378:metabolism; D001861:Bone Regeneration / Q000502:physiology; D001842:Bone and Bones / Q000166:cytology / Q000473:pathology / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D004548:Elasticity; D015870:Gene Expression; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry* / Q000378:metabolism; D020100:Hydrogels / Q000737:chemistry / Q000378:metabolism; D008297:Male; D008422:Materials Testing; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology; D008970:Molecular Weight; D051381:Rats; D017207:Rats, Sprague-Dawley; D023822:Tissue Engineering / Q000379:methods*; D014783:Viscosity
D001672:Biocompatible Materials; D020100:Hydrogels; D006820:Hyaluronic Acid; D020782:Matrix Metalloproteinases