Platelet lysate incorporating with alginate sodium- chondroitin sulfate hydrogel improve mechanical properties and enhances chondrocyte differentiation
,1 David eglin
,2 Mauro alini
,3 Ali samadi kochaksaraei
,4 Mohammadreza baghaban eslaminejad
,5 Majid safa
1. Department of Tissue engineering & Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
2. AO Research Institute Davos, Davos Platz, Switzerland
3. AO Research Institute Davos, Davos Platz, Switzerland
4. Department of Tissue engineering & Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
5. Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
6. Department of Hematology, Iran University of medical science, Associate Professor of Hematology. Tehran/ Iran
Platelet lysate (pl) is a rich component in growth factors and anti-inflammatory cytokines can improve chondrocyte differentiation, but mechanically unstable, in contrast, alginate (alg) is a natural polysaccharide exhibiting excellent biocompatibility and biodegradability that use for a broad range of applications as a biomaterial for tissue repair and regeneration such as articular cartilage regeneration, yet there are no enough intrinsic biological signals. in another hand chondroitin sulfate (cs) as an extracellular matrix of cartilage enhance chondrogenicity and mechanical properties of hydrogel. we hypothesized that the advantage of this system maybe combined in one hydrogel which can be easy translated into clinical setting.
The three parts hydrogel have been prepared with combination of alg (1.5%w/v), cs(1%w/v) and pl(20%v/v) and analyzed in terms of mechanical compression and rheometer properties. the cellular behavior on hydrogel was evaluated with live/dead assay, and chondrogenicity was subsequently evaluated under in vitro conditions by real-time pcr, gag assay and collagen content assay. we have done all experiment in 4 different groups: 1- alg, 2- alg-cs, 3- alg-pl, and 4- alg-cs-pl.
The result of mechanical analysis of hydrogels confirmed higher mechanical strength of alg-cs-pl hydrogel. live/dead assay demonstrated the viability of hbm-mscs in the four different hydrogels. chondrogenic activity of hbm-mscs that included collagen and gag content significantly increased in the alg-cs-pl hydrogel. real-time pcr also confirmed alg-cs-pl hydrogel increase expression of chondrogenic genes such as coll2, aggrecan, and decrease coll 10, alp,and coll1 compare to another group.
Therefore, alginate hydrogel that contained pl and cs promoted chondrogenic differentiation of human-bone marrow derived mesenchymal stem cells.
Platelet lysate, chondrogenesis, mesenchymal stem cells, tissue engineering