Effect of Muscle Extract and Graphene Oxide on Muscle Structure of Chicken Embryos

SIMPLE SUMMARY: Genetic selection of broilers increased muscle growth; however, very fast growth can lead to pathological conditions caused by the deficiency of nutrients. The number of muscle cells is mainly formed during the embryonic period, and consequently, in ovo supplementation of proteins to embryos may impact future muscle structure. We hypothesized that proteins from chicken embryo muscle extract (CEME) caused by the unique, natural composition and biocompatibility can supply additional proteins. However, supplemented proteins are actively metabolized, which may reduce their utilization for improved muscle synthesis. Nevertheless, CEME can be transported and protected by graphene oxide (GO). The objective of the present work was to investigate the effects of in ovo-injected CEME and the complex of GO-CEME on embryonic cell cultures and the growth of chicken embryo hind limb muscle. Toxicity and cell proliferation were measured in vitro with cell cultures and mortality, morphology, histology, and blood biochemistry in vivo with embryos. CEME increased the number of cells and nuclei in muscle, but the complex GO-CEME did not further improve the muscle structure. The results indicate a vital role of CEME as in ovo enhancer of muscle development in broilers. ABSTRACT: The effects of CEME and it complex with GO injected in ovo on the growth and development of chicken embryo hindlimb muscle were investigated. First, the preliminary in vitro study on primary muscle precursor cell culture obtained from a nine-day-old chicken embryo was performed to assess toxicity (MTT assay) of CEME, GO (100 ppm) and it complex with different concentrations (1, 2, 5, and 10 wt.%). The effect on cell proliferation was investigated by BrdU assay. CEME at concentrations 1–5% increased cell proliferation, but not the complex with GO. In vitro cytotoxicity was highest in 10% and GO groups. Next, the main experiment with chicken embryos was performed with CEME, GO and it complex injected in ovo on day one of embryogenesis. On day 20 of embryogenesis survival, morphological development, histological structure of the muscle, and biochemical parameters of blood serum of the embryos were measured. No negative effect on mortality, body weight, or biochemistry of blood after use of CEME or GO-CEME complexes was observed. Interestingly, the slight toxicity of GO, observed in in vitro studies, was not observed in vivo. The use of CEME at the levels of 2% and 5% improved the structure of the lower limb muscle by increasing the number of cells, and the administration of 2% CEME increased the number of nuclei visible in the stained cross-section of the muscle. The complex GO-CEME did not further improve the muscle structure. The results indicate that CEME can be applied as an in ovo enhancer of muscle development in broilers.