K.V. Кulakova, S.N. Bugrov, D.Ya. Aleyinik, A.A. Struchkov

Different collagen-based skin substitutes are used for local treatment of wounds of various genesis. The current trend is to provide a combination of bioactive dressings containing regeneration stimulants, including cultured cells, in combination with different antimicrobial agents, antioxidants which are being gradually desorbed into the wound in the required dosage. Within the framework of creating such skin substitute we have developed wound covering, which is a heterogeneous two-layer membrane that thanks to of the nature of its structure may include a wide range of biologically active substances. In its basic version the material, patented by the team of authors, consists of collagen and polysaccharide components. Neutral collagen is released by the original method from xenogeneic dermis. The collagen coating layer may be impregnated with particulate demeneralized xenogenic bone matrix (DBM). The components are selected due to their ability to support the regeneration process. Amorphous dermal collagen type III can build its own collagen fibers while DBM is a structured type I collagen, and thus can be an effective bearer of various biologically active substances. Polysaccharide (agar) coating layer insulates the surface of the wound from the environment and provides the necessary physical properties of the coating (strength, elasticity, adhesion and adsorption, permeability to gases and liquids). Depending on the application, antibacterial agents or cultured cells may be administered into the base material. During the first stage wound healing properties of the wound covering Generation I (DBM / rivanol / gentamycin) were studied in vivo (120 white nonlinear rats) on a model of scalped full-thickness wounds. As a result, it was shown that the developed collagen material leads to acceleration of regeneration, which shortens epithelialisation as compared to the control group (spontaneous healing wounds, differences between the groups are statistically significant.) During the second stage we carried out a preliminary study of the effectiveness of collagen material on the model of burn wounds (white Wistar rats, contact scald burns of up to 2% of the surface area of the body). Positive results were obtained, indicating a reduction in terms of healing of burns in the experiment. Further expansion of applications of our material is thought to be adapting it to its application together with the culture of fibroblasts. A method of gentle ultraviolet sterilization was developed which provides 100% sterility and composition and manufacturing process were modified to enhance strength characteristics. The collagen generation II material was used in cell culture substrate. A series of in vitro experiments to assess the viability of fibroblasts in culture conditions with collagen material. As a result, presence of single cells on the surface of the collagen gel film substrate was demonstrated that after 24 hours and after 48 hours formation of a monolayerof fibroblasts was observed on the surface of the material. The received data so far confirm the effectiveness of our developed biologically active materials based on collagen for treatment of skin wounds and prospects of using their advanced modifications to replace the deeper tissue defects of various origin.