טכניון מכון טכנולוגי לישראל
הטכניון מכון טכנולוגי לישראל - בית הספר ללימודי מוסמכים  
M.Sc Thesis
M.Sc StudentShani Levi Carmit
SubjectUse of Growth Factors, Cytokines and Keratinocytes
Incorporated into Biodegradable Hydrogel for
Enhancement of Wound Healing
DepartmentDepartment of Biotechnology
Supervisor Ms. Erella Livne (Deceased)


Abstract

Wound healing is a complex programmed sequence of cellular and molecular processes including inflammation, cell migration, angiogenesis, provisional matrix synthesis, collagen deposition and reepithelialization. Growth factors and cytokines like activin and IL-8 are considered candidates for therapeutics because they are synthesized by and stimulate cells required for tissue repair and they are deficient in chronic wounds.

Skin autografts are produced by culturing keratinocytes to generate epidermal skin grafts, but the success of these procedures has been variable with graft failure resulting in many cases after promising initial engraftment. Scaffolds containing growth factors and keratinocytes can serve as biological skin grafts.

The purpose of the present study was to design biological scaffold impregnated with growth factors and cytokines. To test the best  glutaraldehyde (GA) concentration for biological scaffold impregnated with IL-8 or activin for treatment of burns wound healing. To culture keratinocytes on biological scaffold for improved wound healing. To investigate the in vitro biodegradation of the biological scaffold.

Results indicated that the optimal GA concentration was 10mM. Hydrogel scaffold incorporated with IL-8 or activin did not enhance burn wound healing but it was shown that it can be used for treatment for healing of burn and wounds since it was found to be biocompatible and biodegradable.

Isolated human keratinocytes were cultured on the hydrogel scaffold and results have shown that these cells could grow on the hydrogel scaffold until confluent in the culture dish. It was found that both   MMP-9 and MMP-2 were present in the culture medium, indicating their synthesis and release by the cells. Thus the cultured cells contributed to active biodegradation of the hydrogel.


 It is concluded that collagen-based hydrogel scaffold can serve as carrier for growth factors and can support in vitro growth of keratinocytes for wound healing.