title: In Vivo Immune Responses of Cross-Linked Electrospun Tilapia Collagen Membrane<sup/>.
abstract: Collagen has been used extensively in tissue engineering applications. However, the source of collagen has been primarily bovine and porcine. In view of the potential risk of zoonotic diseases and religious constraints associated with bovine and porcine collagen, fish collagen was examined as an alternative. The aim of this study is to use tilapia fish collagen to develop a cross-linked electrospun membrane to be used as a barrier membrane in guided bone regeneration. As there is limited data available on the cytotoxicity and immunogenicity of cross-linked tilapia collagen, in vitro and in vivo tests were performed to evaluate this in comparison to the commercially available Bio-Gide®membrane. In this study, collagen was extracted and purified from tilapia skin and electrospun into a nanofibrous membrane. The resultant membrane was cross-linked to obtain a cross-linked electrospun tilapia collagen (CETC) membrane, which was characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), degradation studies, cytotoxicity studies, and cell proliferation studies. The membranes were also implanted subcutaneously in rats and the host immune responses were examined. The DSC data showed that cross-linking increased the denaturation temperature of tilapia collagen to 58.3°C ± 1.4°C. The in vitro tests showed that CETC exhibited no cytotoxicity toward murine fibroblast L929 cells, and culture of murine preosteoblast MC3T3-E1 cells demonstrated better proliferation on CETC as compared to Bio-Gide. When implanted in rats, CETC caused a higher production of interleukin IL-6 at early time points as compared to Bio-Gide, but there was no long-term inflammatory responses after the acute inflammation phase. This finding was supported with histology data, which clearly illustrated that CETC has a decreased inflammatory response comparable to the benchmark control group. In all, this study demonstrated the viability for the use of CETC as a tissue engineering scaffold and provides an insight on the in vivo immune responses toward xenogenic collagen scaffolds.
authors.affiliation: 1 School of Chemical and Biomedical Engineering, Nanyang Technological University , Singapore, Singapore .
authors.forename: Ammar Mansoor
authors.initials: AM
authors.lastname: Hassanbhai
authors.affiliation: 1 School of Chemical and Biomedical Engineering, Nanyang Technological University , Singapore, Singapore .
authors.forename: Chau Sang
authors.initials: CS
authors.lastname: Lau
authors.affiliation: 1 School of Chemical and Biomedical Engineering, Nanyang Technological University , Singapore, Singapore .
authors.forename: Feng
authors.initials: F
authors.lastname: Wen
authors.affiliation: 1 School of Chemical and Biomedical Engineering, Nanyang Technological University , Singapore, Singapore .
authors.forename: Praveena
authors.initials: P
authors.lastname: Jayaraman
authors.affiliation: 2 National Dental Centre Singapore , Singapore, Singapore .|3 Duke-NUS Medical School Singapore , Singapore, Singapore .
authors.forename: Bee Tin
authors.initials: BT
authors.lastname: Goh
authors.affiliation: 2 National Dental Centre Singapore , Singapore, Singapore .|3 Duke-NUS Medical School Singapore , Singapore, Singapore .
authors.forename: Na
authors.initials: N
authors.lastname: Yu
authors.affiliation: 1 School of Chemical and Biomedical Engineering, Nanyang Technological University , Singapore, Singapore .
authors.forename: Swee-Hin
authors.initials: SH
authors.lastname: Teoh
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology*; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D016207:Cytokines / Q000378:metabolism; D055119:Elastic Modulus; D020007:Implants, Experimental; D008297:Male; D008567:Membranes, Artificial; D051379:Mice; D010012:Osteogenesis / Q000187:drug effects; D017207:Rats, Sprague-Dawley; D040521:Subcutaneous Tissue / Q000187:drug effects; D013718:Tensile Strength; D017210:Tilapia / Q000378:metabolism*; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D003432:Cross-Linking Reagents; D016207:Cytokines; D008567:Membranes, Artificial; D003094:Collagen
keywords: collagen; cytokines; electrospun; immune response; tilapia