An in vitro biocompatibility testing system

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The utility model provides an extracorporeal biocompatibility (BC) detecting system comprising a culture device, a temperature device, an injection device and a detection device. By means of combination of the devices, the system is used for establishing a standard program to detect BC or environment poison of biomedical materials.

means mutual adaptation biocompatible biomaterials and cells, tissues and physiological systems. The methods include: extraction, direct contact and indirect contact method. Wherein the extraction and direct contact disadvantage is not truly simulate Biomaterials contact with tissue condition; indirect contact method is the cells were cultured in three-dimensional matrix containing agar or collagen were more able to simulate the body's physiological environment, but the drawback is the need for testing at elevated temperatures or join enzyme to liquefy the matrix, resulting in damage and affect the reliability and validity analysis of cells. Since the polyether polyol F127 (PluiOnic F127) are FDA approved for use in the polymer medicine, its solution has a unique thermo-reversible gelation phenomenon, when the concentration of 15-20% (w / w) temperature to physiological temperature 37 ° C will form a gel, and reduced to a liquid at 4 ° C, its other advantage is that no additional crosslinking agent. The utility model is the use of this colloid in a simple and accurate measurement methods to enhance the efficiency and the wider use of in vitro biocompatibility analysis and biocompatibility replace traditional detection methods.

The utility model is an in vitro biocompatibility testing system, comprising: a culture device having an outer layer and an inner layer laminated, the outer layer is a microplate, for placing the sandwich, the sandwich of dish, for placing the inner layer and the inner layer is temperature-dependent aspect of colloid, which is used to give nutrients to the subject and the subject growth; temperature device, which features state-based control sample colloid; injection device The colloid was injected into the sandwich culture systems; and inspection means in the inner test culture apparatus for the growth of test specimen.

This utility model in vitro biocompatibility testing system, the regulation of the temperature range of the temperature of the device is (TC -25 ° c. And culture colloid layer of the device configuration requires maintaining at 0 ° C -4 ° C, and in After configuration subject to sterilization, and then placed inside the injection device. In addition, the culture apparatus further comprises colloidal, cell membrane stabilizers, are Baker's Basal Medium (Dulbecco 's Modified Eagle Medium) and fetal bovine serum (FBS). The colloid Wu is a polyether polyol F127 (Pluronic F127), and the cell membrane stabilizer is corticosterone, tocopherol, coenzyme Q or lecithin, choline.

This utility model in vitro biocompatibility testing system, the test apparatus system detects the subject survival, the test method is 2-methyl mercaptan diphenyl tetrazolium bromine detection (MTT) or trypan blue (Trypan Blue) cell count. And the detection method of the test device (TC _15 ° C liquefied detect which device is provided at the bottom of the sandwich culture pores, the sample liquid can penetrate into the microplate.

The utility model sandwich culture device, which is embedded dish dish.

This utility model in vitro biocompatibility testing system, which may be the subject cells.

This utility model in vitro biocompatibility testing system can be used as biomaterials testing tools.

This utility model in vitro biocompatibility testing system can be used as environmental toxicants detection tools.

The utility of new detection system (101) as shown in Figure 1, includes a culture device (102), a culture device (102) is an enlarged view, please refer to Figure 2, the culture device (102) to trace the outer disk (106), Sandwich embedded dish (107), the inner layer is temperature-dependent aspect colloid (109), which functions as a nutrient to be given specimen (110) and the specimen (110) growth; temperature means (103), Its function is to control colloid (109) aspect. The utility model detection system (101) also includes an injection device (104) as the colloid (109) was injected into the culture dissection device (102); and testing device (105), which means culture apparatus (102) Inside test, test specimen for (110) growth.

The utility model culture device (102) for the preparation of polyether polyols F127 (Pluronic F127) colloidal solution, according to the mixing ratio of DMEM and FBS. In the preparation of the polyether polyol need F127 solution was maintained at 0 ° C_4 ° C, and the solution must first be sterilized, the configuration of the polyether polyol F127 solution was placed in the injection device (104), this step need to 0 ° C-4 ° C, re-use injection device (104), polyether polyols F127 embedded solution was injected into the dish (107), and maintain the temperature of 37 ° C was colloidal state (109), with a polyether polyol F127 After the solution into a Petri dish embedded colloid, the colloid-containing polyether polyol F127 microplate was placed (106).

Example II: a polyether polyol F127 colloid special parts analysis

The utility model is more in 15% (w / w) polyether polyol F127 / are Baker's basal medium with 15% (w / w) polyether polyol F127 / secondary water at different temperatures to viscosity Changes observed polyether polyols F127 gel properties, its preferred embodiment, as shown in Figure 3 is displayed, with the temperature increase, 15% (w / w) polyether polyol F127 / Du Baker's Basal Medium viscosity values ​​are also there is a growing trend, and the strength of colloids between 37~40 ° C reached a peak, and 15% (w / w) polyether 4 yuan alcohol F127 / secondary water viscosity value is not pulled situation.

The utility model specimen (110) directly onto the different concentrations of polyether polyol F127 / are Baker's Basal Medium / fetal bovine serum colloid surface and cultured specimen 24, 36 and 48 hours, and by the testing device (105) Trypan Blue (Trypan Blue) cell count test analysis.

The results are shown in Figure 4, the culture in the specimen 24 hours, due to the osmotic membrane under high concentration of polyether polyol F127 rise while filling the specimen and increased cytotoxicity caused by the survival of a large number of lower specimen; the specimen culture 36 hours, due to the embedded dish having pores (108) so that it has permeability, can reduce the impact of osmotic membrane specimen, polyether polyols F127 concentration 15% (w / w), the inspection can proliferator and maintain 75% specimen viability; its more preferred embodiment, the specimen cultured on polyether polyols F127 in 48 hours, cell survival can still be maintained as high as 80%.

The utility model and the other a better embodiment, as FIG. 8, the specimen embedded in polyether polyol / FBS colloids can be observed cell microencapsulation, the colloid promote inter-specimen and specimen began to gather. Also, the utility model is subject to the preferred embodiment of the cell.

Another embodiment of the utility model as the polyether polyols F127 concentration in 14% (w / w), 15% (w / w) and 16% (w / w), add the cell membrane stabilizer, which is corticosterone (hydrocortisone), coenzyme Q (QlO) and tocopherols, and the specimen cultured on polyether polyols F127 colloid, by the trypan blue cytometry analysis, 5, 6 and 7 show, check cultured for 24 hours, polyether polyols F127 concentration in 14% (w / w)> 15% (w / w) and 16% (w / w), to add to the preferred embodiment of corticosterone concentrations were O. 6 μ Μ, 6 μ M and 60 μ M; concentrations of coenzyme Q I μ Μ, 10 μ M and 100 μ M; tocopherol concentrations were O. 635mM, 6 35mM, 63 5mM and 635mM... And then through the analysis sample toxicity, but because of the embedded the dish with the 0.4μπι micropores (108) so as to have a semi-permeability, therefore the utility model, the sample (test solution) added directly to the culture device (102) and ways to penetrate into the culture within the system, and with the test specimen viability and morphology, known specimen Have cytotoxicity. In addition, the detection method shall be the test apparatus(105) detects 0°C -15°C.

Therefore, the preferred embodiment of the utility model is when you add cell membrane stabilizer may promote cell attachment and proliferation and survival of the subject to maintain higher than controls. And more preferably the concentration of cases in which the cell membrane stabilizer is implemented as corticosterone concentration of 60 μ Μ, coenzyme Q concentration of 100 μ Μ, tocopherol concentration of 635mM.

A person familiar with the art in this field can quickly appreciate the utility model can easily achieve their goals and get the results and advantages mentioned, as well as those which exist in things. The utility model in an in vitro biocompatibility testing system is representative of the preferred embodiment, which is exemplary and is not limited to the utility model in the field. Familiar with the art that modifications will occur in which the Department and other purposes. These modifications are inherent in the spirit of the utility model and define the scope of the patent.

This utility model describes the content and embodiments are disclosed in detail, such that any person skilled in the art can make and use this creation, even if there are at the various alternatives, modifications, and progress of, should still be considered without departing from the spirit and scope of the utility model. All patents and publications mentioned in the specification, and the creation of related fields are in general skills prevail. All patents and publications are incorporated herein by reference to the same extent as if each individual publication were specifically and individually indicated to be incorporated by reference. The invention herein illustrated as appropriate, may be in the absence of any element, or a plurality of elements, limitation or limitations are not specific for the next case in the embodiment disclosed herein. Terms and expression used is as described in the specification and not of limitation, and there is no intent to exclude any use of such equivalent features shown and described or portions thereof and expression of the term, but need to recognize that, in this there may be various changes within the scope of the patent application creation. Therefore, we should understand that although the present invention will be specifically disclosed in accordance with the characteristics of the preferred embodiment and arbitrary, but known to the art that modifications and changes will continue to reveal the contents therein, modifications and variations of the present invention still like within the scope of patent applications.