Composite cell support membranes based on collagen and polycaprolactone for tissue engineering of skin.

The preparation and characterisation of collagen:PCL composites for manufacture of tissue engineered skin substitutes and models are reported. Films having collagen:PCL (w/w) ratios of 1:4, 1:8 and 1:20 were prepared by impregnation of lyophilised collagen mats by PCL solutions followed by solvent evaporation. In vitro assays of collagen release and residual collagen content revealed an expected inverse relationship between the collagen release rate and the content of synthetic polymer in the composite that may be exploited for controlled presentation and release of biopharmaceuticals such as growth factors. DSC analysis revealed the characteristic melting point of PCL at around 60 degrees C and a tendency for the collagen component, at high loading, to impede crystallinity development within the PCL phase. The preparation of fibroblast/composite constructs was investigated using cell culture as a first stage in mimicking the dermal/epidermal structure of skin. Fibroblasts were found to attach and proliferate on all the composites investigated reaching a maximum of 2 x 10(5)/cm(2) on 1:20 collagen:PCL materials at day 8 with cell numbers declining thereafter. Keratinocyte growth rates were similar on all types of collagen:PCL materials investigated reaching a maximum of 6.6 x 10(4)/cm(2) at day 6. The results revealed that composite films of collagen and PCL are favourable substrates for growth of fibroblasts and keratinocytes and may find utility for skin repair.

[1]  H Green,et al.  Growth of cultured human epidermal cells into multiple epithelia suitable for grafting. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Jeffrey A. Hubbell,et al.  Biomaterials in Tissue Engineering , 1995, Bio/Technology.

[3]  J. Ricotta,et al.  Use of Dermagraft, a Cultured Human Dermis, to Treat Diabetic Foot Ulcers , 1996, Diabetes Care.

[4]  E Bell,et al.  Living tissue formed in vitro and accepted as skin-equivalent tissue of full thickness. , 1981, Science.

[5]  I. Leigh,et al.  Epidermal cytokines and their roles in cutaneous wound healing , 1991, The British journal of dermatology.

[6]  N. Parenteau,et al.  Development of a stratum corneum and barrier function in an organotypic skin culture , 2004, Archives of Dermatological Research.

[7]  S. Boyce,et al.  Burn wound closure with cultured autologous keratinocytes and fibroblasts attached to a collagen-glycosaminoglycan substrate. , 1989, JAMA.

[8]  Paul Martin,et al.  Wound Healing--Aiming for Perfect Skin Regeneration , 1997, Science.

[9]  I. Yannas.  Studies on the biological activity of the dermal regeneration template , 1998, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[10]  J. Hansbrough,et al.  Clinical trials of a living dermal tissue replacement placed beneath meshed, split-thickness skin grafts on excised burn wounds. , 1992, The Journal of burn care & rehabilitation.

[11]  G. Vunjak‐Novakovic,et al.  Culture of organized cell communities. , 1998, Advanced drug delivery reviews.

[12]  M. Pierschbacher,et al.  Enhancement of cell interactions with collagen/glycosaminoglycan matrices by RGD derivatization. , 1997, Biomaterials.

[13]  A. Coombes,et al.  Biocomposites of non-crosslinked natural and synthetic polymers. , 2002, Biomaterials.

[14]  Richard O. Hynes,et al.  Integrins: Versatility, modulation, and signaling in cell adhesion , 1992, Cell.

[15]  K. Athanasiou,et al.  Extracellular matrix cell adhesion peptides: functional applications in orthopedic materials. , 2000, Tissue engineering.

[16]  I. Yannas,et al.  Wound tissue can utilize a polymeric template to synthesize a functional extension of skin. , 1982, Science.

[17]  Y. Tokiwa,et al.  Hydrolysis of polycaprolactone fibers by lipase: Effects of draw ratio on enzymatic degradation , 1995 .

[18]  M. P. Welch,et al.  Temporal relationships of F-actin bundle formation, collagen and fibronectin matrix assembly, and fibronectin receptor expression to wound contraction , 1990, The Journal of cell biology.

[19]  Richard A.F. Clark,et al.  The Molecular and Cellular Biology of Wound Repair , 2012, Springer US.

[20]  F. Silver,et al.  Fibroblast growth on a porous collagen sponge containing hyaluronic acid and fibronectin. , 1987, Biomaterials.

[21]  S. Barttelbort,et al.  Composite autologous-allogeneic skin replacement: development and clinical application. , 1987, Plastic and reconstructive surgery.

[22]  A. Schindler,et al.  Aliphatic polyesters. I. The degradation of poly(ϵ‐caprolactone) in vivo , 1981 .