Introduction: Cholecyst derived scaffold (CDS) is a biomaterial that has found a variety of application in the regenerative medicine[1]. It is known to induce faster healing of full thickness skin excision wound in animal models[2][3]. This paper compares various physical properties and wound healing potential of CDS with other extrcellular matrix scaffolds of similar origin (such as jejunum derived scaffolds and urinary bladder derived scaffolds) for skin tissue engineering and cutaneous wound healing application.
Materials and Method: Qualitative detection of linker proteins such as collagen IV, collagen VII and laminin in all the scaffolds were determined using immunoflourescence. The proliferation of HaCaT cells and dermal fibroblast on various scaffolds were evaluated by MTT assay. Immunohistochemical staining of skin tissue engineered construct prepared by seeding HaCaT cells on respective scaffolds were performed for PCNA, involucrin and Vitamin D receptor. The physical properties of cholecyst derived scaffold for skin graft application were compared with other extra cellular matrix scaffolds. Wound healing reaction of CDS was compared with other scaffolds on full thickness excision wounds in rabbit model (after approval by the institutional ethics committee). In vivo wound healing experiment was also performed on third degree burn wounds using cholecyst derived scaffold and autologous fibroblast loaded cholecyst derived scaffold. Healing reaction of the damaged skin tissue was quantified by histomorphometry and immunohistomorphometry techniques in sections of histology and immunohistochemistry stained slides to access the progress of the healing process.
Results and Discussion: Collagen IV, collagen VII and laminin were detected in all the scaffolds. In this study, a model skin tissue constructs was fabricated by seeding HaCaT cells on cholecyst derived scaffold jejunum derived scaffold and urinary bladder scaffolds originally prepared by a a non-detergent/nonenzymatic method[2] and the immunohistochemical analysis of this skin tissue constructs showed that the HaCaT cells were proliferating and functionally active on the cholecyst derived scaffold. Wound healing in vivo experiment using cholecyst derived scaffold as skin graft in comparison to other skin grafts showed that CDS aided speedy wound healing by granulation tissue formation and minimal fibrosis. Loading of fibroblast cells on CDS also aided fast healing of full thickness burn wounds in rabbits.
Conclusion: It can be concluded that cholecyst derived scaffold is a potential biomaterial for fabricating bioartificial skin graft for wound healing application.
Govt of India, Dept of Biotechnology for project BT/PR15461/MED/32/16/7/2011 dated 16 Jan 2012,; Govt of India, CSIR for Senior Research fellowship
References:
[1] Burugapalli K, Chan JC, Kelly JL, Pandit A (2008) Buttressing staples with cholecyst-derived extracellular matrix (CEM) reinforces staple lines in an ex vivo peristaltic inflation model. Obes Surg 18:1418-23.
[2] Anilkumar TV, Vineetha VP, Revi D, Muhamed J, Rajan A (2014) Biomaterial properties of cholecyst-derived scaffold recovered by a non-detergent/enzymatic method. J Biomed Mater Res B Appl Biomater 102:1506-16.
[3] Revi D, Vineetha VP, Muhamed J, Rajan A , Anilkumar TV (2013) Porcine cholecyst-derived scaffold promotes full-thickness wound healing in rabbit. J Tissue Eng 4:2041731413518060.