Preadipocyte-loaded collagen scaffolds with enlarged pore size for improved soft tissue engineering.

Extended soft tissue defects after extensive deep burns or tumor resections are still an unresolved problem in plastic and reconstructive surgery. There is a clinical need for an adequate solution to this problem but currently, no adequate implant material is available for the correction of these defects. Since the autologous transplantation of mature adipose tissue gives poor results, this study explores the advantages of using human preadipocytes in collagen sponges for tissue reconstruction purposes. Human preadipocytes of young adults were isolated, cultured, seeded onto collagen sponges with uniform pore size, and implanted into immunodeficient mice. After 24 hours of incubation in vitro and after explantation at 3, 8, and 12 weeks, sponges were examined for macroscopic appearance, weight, thickness, histology, immuno-histochemistry, and ultrastructure. We find good penetration of cells into the scaffold, layers of adipose tissue, and new vessels on all grafts while controls appear unchanged. These results are promising for improving the reconstruction of soft tissue defects.

Preparation of collagen scaffolds and their applications in tissue engineering.

Freeze-dried collagen scaffolds can be used for a variety of medical tissue engineering applications. The pore structure of the scaffolds might play a decisive role for the inoculation, growth and differentiation of the cells. For a controlled 3D-cell growth the pore structure needs to be homogeneous and the pore size individually adjustable. For lyophilised scaffolds, the pore structure is determined by the ice crystal morphology during freezing under steady conditions. Scaffolds with a homogeneous pore structure and a range of pore size between 25 and 100 microns were reached. Cells such as preadipocytes, keratinocytes, and fibroblasts showed to adhere well to the collagen matrix.

Magnification of the pore size in biodegradable collagen sponges.

In tissue engineering cells are often combined with a carrying structure with collagen being a suitable material to form a 3D-scaffold. A process to manufacture collagen sponges with an adjustable and homogeneous structure has been developed at the Helmholtz-Institute. Using this process, collagen suspensions are frozen directionally and subsequently vacuum-dried. One clinical application in which these scaffolds can be used is soft tissue reconstruction. Various soft tissue defects require an adequate replacement, e.g. in the case of severe burn wounds, or after tumour resections. Collagen (type I) sponges, which are cultured with preadipocytes, may be used to regenerate such defects. In this case, pore sizes of approximately 100 microm are desired to allow a complete differentiation of preadipocytes into adipocytes. Based on known technology to manufacture collagen sponges with an adjustable and homogeneous pore structure, research on the increase of pore size beyond the previous limit of 40 microm was necessary in order to enable soft tissue replacement. A scaffold with an average pore size of 100 microm was obtained.