Experimental Study On Biocompatibility Of Human Amniotic Epithelial Cell With Muscle Basal Lamina

The impaired diseases of nervous system result in difficulties of axonal regeneration, leading to nonvolatile loss of motor function and sensory function. Glial scar and cavitates formation are the main obstacle for axonal regeneration in the spinal cord injury. It is important to bridge injured spinal cord by a graft crossing glial scar and cavitates. In recent years, tissue engineering brings new desire for treating spinal cord injury. Scafford not only bridges injured spinal cord, but also charges tissues,cells and neurotrophic factors. In addition, it can be made into specified shape to conduct axonal directional growth.The muscle basal lamina has a good biocompatibility which can promote cell adherent and proliferation. Also it has low immunogenicity and its degradation products are avirulent. In addition it can be made into different shape according to the requirement. The basal lamina itself represents a component of the extracellular matrix and the basal lamina tubes provide spaces for the regenerating axons to grow through. Internal structure of muscle basal lamina is similar to endoneural membrane. Thus, regenerative nerve fibers grow better ordered and reasonable. In a conclusion, basal lamina tubes, as in acellular muscles, represent favorable scaffolds in tissue engineering.It has been reported that human amniotic epithelial cells (HAECs) deliver neurotrophic factors such as BDNF,NT-3. HAECs provide microenvironment for host neuronal growth. Also, HAECs have low immunogenicity, convenient materials, abundant sources, not ethical and legal aspects. Therefore HAECs are ideal seed cells.Object To investigate the biocompatibility of the muscle basal lamina with human amniotic epithelial cell. Thus, people can apply nervous tissue engineering to treat the impaired diseases of nervous system.Methods The muscle basal lamina was manufactured by chemical extraction. Whether or not the muscular cells were removed could be detected by regular histological observation. Extracellular matrix in the muscle basal lamina was detected by VG+ET staining. Human amniotic epithelial cells (HAECs) were cultured in vitro. HAECs were seeded in the muscle basal lamina at a density of 2×106/ml in vitro when they were in a good condition. The cultivated HAECs in the muscle basal lamina were examined by scanning electron microscope. Brain-derived neurotrophic factor (BDNF), neurotrophic-3 (NT-3) and BrdU were detected in the cultivated HAECs in the muscle basal lamina by immunohistochemistry.Resluts By gross observation, the muscle presents rosiness, which is opaque and rigid. The muscle basal lamina presents ivory, semitransparent and soft by chemical extraction. There is little change in shape after chemical process. In longitudinal section of the muscle basal lamina by HE staining, the cell compoments completely disappear after the muscle has been extracted. The scaffold of acellular muscle is arranged in parallel tubules composed of collagen fibers and elastic fibers which are left intact. HAECs generate well in the muscle basal lamina and distribute uniformity in vitro. BrdU positive cells which are detected by immunohistochemistry show that HAECs in the muscle basal lamina can proliferate. NT-3 and BDNF positive cells are detected in the cytoplasm by immunohistochemistry. By scanning electron microscope acellular muscle is arranged in parallel tubules and HAECs generate well in the muscle basal lamina. HAECs present sphere and short prominence. Conclusion The muscle basal lamina is manufactured successfully according to requirement of nervous tissue engineering scaffold. HAECs generate well in the muscle basal lamina in vitro and synthesize NT-3 and BDNF.