Experimental Report on the Selection and Use of Biomedicals in the Field of tissue Engineering#Experimental report on the selection and application of biological materials in tissue engineering
** I. Introduction **
The purpose of tissue engineering is to repair or regenerate damaged tissues by combining biological materials, cells, and biological active factors. Biomedicals played a crucial role in this process, and their selection directly affected the success or failure of tissue engineering. The purpose of this experiment was to explore the basis for the selection of biological materials and their application in tissue engineering.
** 2. Biological Material's classification and characteristics **
(I) Inert biological materials
1. ** Medical metal material **
- include stainless steel, titanium alloy, Cobalt base alloy, nickel-titanium alloy, silver-mercury alloy, and that like. This type of material has good mechanical properties, such as high strength and toughness, and can maintain a relatively stable structure in the biological environment without or with only weak chemical reactions. Clinically, it could be used to make artificial joints and other implanted devices that needed to withstand large mechanical forces.
2. ** Medical non-metallic material **
- For example, ceramic materials such as aluminum dioxide, zirconium dioxide, titanium dioxide, silicon dioxide, magnesium-dioxide, and calcium chlorite-acid. They had good compatibility and high hardness, and could be used to make artificial bones and other repair materials.
3. ** Medical high molecular material **
- There were many varieties, such as PE, PG, PVP, Pan, PM, PUR, Si rubber, PVP fiber, carbon fiber, etc. Its advantage lay in its strong machinability, which could be made into medical devices or tissue engineering matrices of various shapes and structures, and some of the high molecular materials had good flexibility.
4. ** Medical composite material **
- It was made of two or more materials with different chemical properties. For example, fiber reinforced plastic and metal-ceramic composite materials. The composite material could combine the advantages of different materials, such as combining the machinability of a high molecular material with the high strength of a metal or ceramic to meet the needs of different tissue engineering.
(2) Bioactive materials
1. ** Bioactive metals and alloys **
- Able to interact with living organisms and have specific functions. Its physical form, topography, or size can be specially designed or designed for its function.
2. ** Bioactive Inorganic Matter **
- Including ceramic, glass, and carbon-based materials. These materials could form chemical bonds with biological tissues through specific surface treatments or structural designs to promote tissue repair and regeneration.
3. ** Bioactive Polymers and Gels **
- It can be used to load cells and transfer growth factors. Its soft texture and controllable physical and chemical properties are conducive to cell attachment, reproduction, and differentiation.
4. ** Natural Bioactive Material **
- It has good biological compatibility and biological activity, and can be obtained from a wide range of sources. It can be extracted from living organisms or obtained through biochemistry.
5. ** Bioactive composite material for human or animal use **
- For example, an implant, a tissue engineering stent, a cell/drug/gene carrier, an imaging and sensing device, etc. These composite materials could combine bio-active materials with other functional materials to achieve the integration of multiple functions.
** 3. The basis for the selection of biological materials **
(I) Biocompatibility
1. The biological material should not cause immune reaction in the body, be non-invasive, not teratogenic, not cancerous, and not cause adverse reactions in the body, such as blood clot, hemolation, and chemotherapy.
2. The surface properties of the material had a great impact on the compatibility, such as the surface toughness, chemical active groups, and so on. Materials with smooth surfaces and suitable chemical active groups were more conducive to cell attachment and growth.
(II) Mechanical properties
1. Depending on the target tissue of tissue engineering, the requirements for the mechanical properties of the biological materials were different. For example, materials used for bone tissue engineering needed to have high compression strength and elasticity to withstand the mechanical load of the bone, while materials used for soft tissue engineering needed to have good flexibility and elasticity.
2. The mechanical properties of the material should also match the growth and repair process of the tissue. In the process of tissue regeneration, the mechanical properties of the material may change with the growth of the tissue. It was necessary to ensure that the material could provide sufficient support throughout the entire process without hindering the growth of the tissue.
(3) Biodegrading
1. For some tissue engineering applications, such as the use of sutures and bone repair matrices, the materials needed to be sufficiently biodegraded. The rate of decomposition should be coordinated with the rate of tissue regeneration. Too fast decomposition may lead to incomplete tissue repair, while too slow decomposition may affect the normal functional recovery of the tissue.
2. Biodegraded products should be non-toxic and can be eliminated by the body's metabolism. They should not accumulate in the body and cause damage to the body.
(IV) Porosity and Microstructure
1. A suitable void ratio would facilitate the migration of cells, the exchange of nutrients, and the discharge of waste products. Higher porosities could provide more space for cells to grow, but at the same time, it would affect the mechanical properties of the material. A balance between the two was needed.
2. Microstructures such as fibers and pores could also affect the behavior of cells. For example, the fiber structure could mimic the structure of the matrix, which was beneficial for the directional growth of cells.
** 4. Experiment on the application of biological materials **
(I) Experiment Purpose
Testing the application effect of the selected materials in tissue engineering, including cell attachment, reproduction, and differentiation, as well as the material's compatibility and biochemistry.
(2) Experimental Materials
1. The medical polylactic-co-gly colic acid (Plga) was selected as the bio-degrading high molecular material, which had good biological compatibility and could adjust the rate of decomposition.
2. As the seed cells, the bone cells were used to simulate the cell behavior in bone tissue engineering.
(3) Experimental Method
1. material preparation
- The pore size and the porosity of the matrix could be adjusted by controlling the preparation process.
2. cell culture
- The bone cells were seeded onto the PDBG stent and cultured under suitable cell culture conditions (such as 37°C, 5% CO2).
3. test index
- Cell attachment: Observe the attachment of cells on the surface of the stent through a scanning electron microscope, and calculate the number and shape of the attached cells.
- Cell proliferations: Cell counting kit (CCK - 8) was used to detect the cell proliferations at different time points (such as 1, 3, 5, and 7 days), and the cell proliferations curve was drawn.
- Cell differentiation: To detect the markers related to the differentiation of the bone blasts (such as the activity of Alkaline Phosphatase, the content of Bone Galexin, etc.), and to evaluate the degree of cell differentiation on the PLGA stent.
- Biocompatibility: implant the cell-seeded stent into the animal body (such as the mouse's skin), and observe the tissue reaction at different time points (such as 1, 2, and 4 weeks), including the degree of inflammation and the formation of blood vessels in the tissues around the material.
- Biodegrade: The rate of the biodegrade of the PLGA stent was evaluated by measuring the mass loss and molecular weight change of the material in the simulated environment in the body or in the body.
(4) Experimental results
1. cell adhesion
- The results of the scanning electron microscope showed that the bone cells could adhere to the surface of the PLGA matrix well, and the cells extended pseudopodia to interact with the surface of the matrix. As the culture time was prolonged, the number of adhered cells gradually increased.
2. cell proliferation
- The CCK - 8 test results showed that the cell proliferations showed a gradual upward trend in the first 7 days of culture, indicating that the PLGA stent had no inhibition effect on the proliferations of the bone cells.
3. cell differentiation
- The results of Alkaline Phosphatase Activity and Bone Calcinin content test showed that the bone cells could differentiate normally on the PLGA matrix. As the culture time increased, the expression level of the differenciation-related markers gradually increased.
4. BC (biocompatibility)
- In the animal implant experiment, a slight inflammation reaction was observed at 1 week, and the inflammation reaction gradually reduced after 2 weeks. At 4 weeks, there was obvious blood vessel formation in the tissue around the material, indicating that the PLGA stent had good compatibility.
5. biodegradability of
- The results of the in vitro-simulation experiment showed that the PLGA stent gradually degraded within a certain period of time (such as 8 - 12 weeks). The mass loss and molecular weight reduction met the expected decomposition curve, and the decomposition products did not have any adverse effects on the surrounding environment.
** 5. conclusion **
1. In tissue engineering, the selection of materials required comprehensive consideration of many factors, such as the biological compatibility, mechanical properties, biodegrade, and the micro-structure.
2. Through the application experiment in bone tissue engineering, it was proved that the material had good cell attachment, proliferating and differentiated support ability, as well as good compatibility and biodegrading. It was an ideal biological material for bone tissue engineering. However, different tissue engineering applications may require further optimization of the performance of the materials to meet specific tissue repair and regeneration needs. Future research needed to explore the performance and application potential of more types of biological materials, as well as develop new materials preparation techniques and surface modification methods to improve the effect of tissue engineering.
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