T/CSBM 0050.3-2024 Evaluation of Wound Repair Material Efficacy - Part 3: Evaluation Method for Stimulation Factor Shielding
T/CSBM 0050.3-2024 Evaluation of Wound Repair Material Efficacy - Part 3: Evaluation Method for Stimulation Factor Shielding
Basic Information
Scope
Scope: This document specifies the determination of stimulus concentration in vitro evaluation of wound repair materials, the physical barrier cell viability test, and the physical barrier cell migration test. This document is applicable to hydrogels or solid wound repair biomaterials for protecting the viability and migration of wound cells; Main technical content: 4. Stimulus concentration determination test 4.1 Principle: The stimulus is dissolved in the culture medium to prepare samples with different graded concentrations. The cytotoxicity of the stimulus at different concentrations is determined according to GB/T 16886.5 to determine the survival rates of 40%, 50%, and 60% of the stimulus. 4.2 Instruments, reagents, and consumables, cell lines, stimuli 4.2.1 Instruments: (a) CO2 cell culture incubator; (b) constant temperature water bath shaker; (c) high-pressure steam sterilizer; (d) inverted fluorescence microscope; (e) cell counter; (f) constant temperature water bath shaker; (g) enzyme analyzer; (h) electronic balance; (i) liquid nitrogen tank; (j) freezer centrifuge; (k) 96-well plate; (l) cell culture flask or cell culture dish. 4.2.2 Reagents and consumables: (a) DMEM low-sugar medium containing 10% FBS; (b) dimethyl sulfoxide (DMSO); (c) isopropanol; (d) high-density polyethylene. 4.2.3 Cell lines 4.2.3.1 Cell lines available for the test include: (a) fibroblasts; (b) skin epithelial cells; (c) esophageal epithelial cells; (d) gastric mucosal epithelial cells; (e) intestinal mucosal epithelial cells. 4.2.3.2 For repair materials used in different parts, select appropriate cell lines, such as skin repair materials can use skin cells, and esophageal repair materials can select esophageal epithelial cells. 4.4 Test method: The MTT cytotoxicity test method specified in Appendix C of GB/T 16886.5-2017 is used. The processing conditions, detection model, and observation time are according to the table.