A novel uniform strain chamber for adherent cell loading devices

Abstract

The present invention discloses a novel uniform strain chamber for an adherent cell loading device, which relates to the field of cell mechanics. Cells are planted in the novel uniform strain chamber of the adherent cell loading device, and the four positioning holes of the chamber are placed on a fixed pile. The fixed pile is connected to the stator. The controller controls the stepper motor, and the rotational motion of the motor is converted into linear motion through a transmission screw and transmitted to the mover, thereby indirectly strain stimulating the cells in the chamber cavity, thereby realizing relevant mechanical biology research on the cells. The strain of the adherent cell chamber in the existing cell tensile loading device is uneven, and the experimental results have large errors. In the invention, the elastic basement membrane of the chamber is designed to have one side as a plane, i.e., the part where the planted cells are poured into the cell culture medium, and the other side as a curved surface, i.e., an optimized surface. The longitudinal cross-section is an M-shaped structure, and its thickness variation meets the uniformity requirement of the strain of the plane part of the basement membrane during the chamber loading process, thereby increasing the uniform area of ​​the adherent cells in the strain loading, and therefore has high use value.

Description

Technical field:

[0001] The patent of this invention is a novel uniform strain chamber for an adherent cell loading device, which relates to the field of cell mechanics technology. Background technology:

[0002] Cell mechanics is a cutting-edge field in modern biomechanics. Understanding the impact of mechanical factors on cell behavior is fundamental and crucial for finding the optimal method for loading cells. After years of refinement and effort, researchers have developed a variety of cell loading instruments to suit diverse experimental needs.

[0003] Based on the mechanical environment of bone-related cells, the bioreactor that applies mechanical stimulation to cells in vitro is an experimental device for periodic tensile strain of cells, which is expected to provide a suitable mechanical environment for the in vitro culture of bone-related cells and provide an experimental basis for studying the molecular mechanisms of cells. The adherent cell strain loading instrument is based on substrate deformation loading technology. Through the control system, the rotation speed and rotation direction of the stepper motor are controlled, and different tensile strains and tensile frequencies are applied to the elastic basement membrane to simulate the mechanical stimulation of cells in vivo. Studying the response of cells to mechanical stimulation of different frequencies and strains and discovering the mechanism of the influence of mechanical stimulation on cells have important theoretical and practical guiding significance.

[0004] Among the current methods for in vitro mechanical loading of cells, substrate deformation loading technology is suitable for in vitro mechanical experiments on various cells. It is not limited by the experimental cycle and is currently an ideal experimental device. However, the prominent problem is that the substrate deformation uniformity of existing flat cell strain loading chambers is low. Summary of the invention:

[0005] The technical problem to be solved by this invention is to design a novel uniform strain chamber for an adherent cell loading device. The structural design of the chamber utilizes a method of continuously adjusting the thickness of the elastic basement membrane. This increases the uniform strain area of ​​the chamber during stretching, providing adherent cells with more uniform mechanical stimulation.

[0006] The technical solution employed by the present invention is to provide a novel uniform strain chamber for an adherent cell loading device. The chamber exhibits different structures at three different stretching rates: 1%, 5%, and 10%. The difference lies in the distance between the upper plane of the elastic basement membrane and the lower curved surface of the elastic basement membrane, i.e., the thickness of the elastic basement membrane. Any longitudinal cross-section of the lower curved surface 4 of the elastic basement membrane is M-shaped. The elastic basement membrane structure with the M-shaped surface can adopt different curved surface structures according to different stretching rates.

[0007] The present invention provides three chamber structures under different stretching rates. The design principle is to increase the thickness of the part with larger strain, and the strain value during the actual loading process will be reduced; reduce the thickness of the part with smaller strain, and the strain value during the actual loading process will be increased. Therefore, the thickness of the elastic basement membrane is different under different stretching rates.

[0008] The beneficial effect of the present invention is that the present invention simulates the actual tensile loading of the cell-seeding chamber in the adherent cell loading device, and changes the thickness of the elastic basement membrane according to the simulation results to improve the strain uniformity of the chamber during the stretching process. The chamber structure obtained according to this method has a maximum strain uniformity of 113% higher than that of the traditional chamber during the stretching process. The improvement in the chamber stretching uniformity greatly increases the accuracy of the device, reduces the error in the experimental process, and makes the experimental results more accurate. Description of the drawings:

[0009] Figure 1 It is a device for fixing a new type of uniform strain chamber. 101 is a static fixing hole, 102 is a fixing pile, and 103 is a dynamic fixing hole.

[0010] Figure 2 It is a three-dimensional model of a novel uniform strain chamber and a fixing device. 201 is the novel uniform strain chamber, and 202 is the device for fixing the novel uniform strain chamber.

[0011] Figure 3 This is a model of the new uniform strain chamber of the adherent cell loading device, a is the front model of the chamber, and b is the bottom model of the chamber.

[0012] Figure 4 is the elastic basement membrane of the novel uniform strain chamber of the adherent cell loading device, c is the upper plane model of the elastic basement membrane, and d is the lower curved surface model of the elastic basement membrane.

[0013] Figure 5 It is a longitudinal cross-section model of an elastic basement membrane with a symmetrical structure. The red part represents the cross section, the number on the left is the distance between adjacent cross sections, and the number on the right is the cross section number.

[0014] Figure 6 This is the shape and thickness of the cross section of the elastic basement membrane when the stretch rate is 1%. Figure 6 Curves 601 to 610 correspond to Figure 5 The cross sections ① to ⑩.

[0015] Figure 7 This is the shape and thickness of the cross section of the elastic basement membrane when the stretch rate is 5%. Figure 7 Curves 701 to 710 correspond to Figure 5 The cross sections ① to ⑩.

[0016] Figure 8 This is the shape and thickness of the cross section of the elastic basement membrane when the stretch rate is 10%. Figure 8 Curves 801 to 810 correspond to Figure 5 The cross sections ① to ⑩.

[0017] Figure 9 Figure 901 shows the strain area ratio for the traditional cell and the new uniform strain cell during stretching at a stretching rate of 1%, while Figure 902 shows the strain area ratio for the new uniform strain cell.

[0018] Figure 10 Figure 1001 shows the strain area ratio for the traditional cell and the new uniform strain cell during stretching at a stretching rate of 5%, while Figure 1002 shows the strain area ratio for the new uniform strain cell.

[0019] Figure 11 Figure 1101 shows the strain area ratio of the traditional cell and the new uniform strain cell during stretching, when the stretching ratio is 10%. Figure 1102 shows the strain area ratio of the new uniform strain cell.

[0020] Figure 12 This is the overall model diagram of the new uniform strain chamber of the adherent cell strain loading device, 1 is the static positioning hole, 2 is the dynamic positioning hole, 3 is the upper end plane of the elastic basement membrane, and 4 is the lower end curved surface of the elastic basement membrane. Specific implementation method:

[0021] Next, in order to make the purpose, method of use and efficacy of the present invention more clearly understood, embodiments of the present invention are described in detail with reference to the accompanying drawings to further illustrate the present invention. The exemplary embodiments of the present invention and their description are only used to explain the present invention and are not intended to limit the present invention.

[0022] The present invention is described in detail with reference to the accompanying drawings and embodiments as follows:

[0023] Reference Figure 1 As shown, 101 and 1033 are respectively the static and dynamic fixing holes for fixing the new uniform strain chamber, and 102 is a fixing pile. Figure 2 As shown in the figure, the static and dynamic positioning holes of the new uniform strain chamber are matched with the static and dynamic fixing holes of the fixing device respectively. The fixed piles support the entire model. During the process of motion loading, Figure 2 The left end of the fixed pile is fixed and stationary, while the right end of the fixed pile is stretched and displaced, thereby mechanically stimulating the cells in the chamber.

[0024] Reference Figure 3As shown, 301 is the front model of the strain loading chamber for adherent cells, and the elastic basement membrane plane of the inner cavity is used to plant cells. The cell culture medium can be poured into the plane to provide a suitable growth environment for the cells, thereby achieving a better stretching effect; 302 is the bottom model of the strain loading chamber for adherent cells, and the lower end of the elastic basement membrane is a curved surface. The curved surface structure has better strain loading area uniformity than the flat structure.

[0025] Reference Figure 4 As shown, 401 is the upper end plane of the elastic basement membrane of the adherent cell strain loading chamber, which is used to plant cells and place cell culture medium. The cells are close to the plane. As the plane is strain loaded and stretched, the cells thereon also receive mechanical tension, completing the mechanical stimulation of the cells; 402 is the lower end curved surface of the elastic basement membrane of the adherent cell strain loading chamber. The traditional flat surface cannot provide a uniform strain area effect. The curved surface can increase the uniformity of the strain area of ​​the chamber during the stretching process, making the experimental results more accurate.

[0026] Reference Figure 5 As shown, Figure 5 This is a cross-sectional model of the elastic basement membrane of the new uniform strain chamber of the adherent cell loading device. Since the elastic basement membrane is a symmetrical structure, the cross-sectional shape along the longitudinal symmetry axis of the elastic basement membrane can be seen to be "M" shaped; the total length of the elastic basement membrane is 40mm, and its half model is 20mm. Figure 5 The numbers on the left are the distances between adjacent sections. 10 sections are divided according to the distances. Figure 5 The numbers ① to ⑩ on the right are the numbers of the 10 sections.

[0027] Reference Figure 6 、 7 , 8 show the longitudinal cross-sectional shape and thickness data of the elastic basement membrane of the adherent cell strain loading chamber when the stretching rates are 1%, 5% and 10%, respectively. Figure 6 、 7 , 8 in the 601, 701, 801 curves correspond to Figure 5 The first section ① in Figure 6 、 7 The coordinate axes of the curves 601, 701, and 801 in 8 represent the longitudinal section lines of the upper plane of the elastic basement membrane. The points on the coordinate axes divide the longitudinal section lines into 28 points. The data of the points are symmetrical. The data of the first 14 points have been marked on the curves, and the data values ​​are the thickness of the elastic basement membrane. The same applies to curves 602~610, 702~710, and 802~810.

[0028] according to Figure 6 、 7, 8, the thickness data can be used to draw 10 curves in the three-dimensional software, which can be connected into a plane through lofting. This plane is the curved surface structure of the lower end of the elastic basement membrane of the chamber.

[0029] Reference Figure 9 As shown, when the stretching rate is 1%, the strain uniform area 901 of the traditional cell accounts for 72% during the stretching process, and the strain uniform area 902 of the new uniform strain cell accounts for 90% during the stretching process; Figure 10 As shown, when the stretching rate is 5%, the strain uniform area 1001 of the traditional cell accounts for 41% during the stretching process, and the strain uniform area 1002 of the new uniform strain cell accounts for 88% during the stretching process; Figure 11 As shown, when the stretching rate is 10%, the strain uniform area 1101 of the traditional chamber accounts for 71% during the stretching process, and the strain uniform area 1102 of the new uniform strain chamber accounts for 88% during the stretching process.

[0030] In summary, the strain uniformity area of ​​the novel uniform strain chamber in the adherent cell loading device of the present invention increased from 72% to 90% at a stretch rate of 1%, from 41% to 88% at a stretch rate of 5%, and from 71% to 88% at a stretch rate of 10%. This improved stretch uniformity of the silicone chamber significantly increases the device's precision, reduces experimental errors, and provides more accurate experimental results.

Claims

1. A novel uniform strain chamber for an adherent cell loading device, characterized by: The chamber structure includes two static positioning holes 1, two dynamic positioning holes 2, an upper end plane 3 of the elastic basement membrane, and a lower end curved surface 4 of the elastic basement membrane; the two static positioning holes 1 are connected to the stator of the fixed pile of the adherent cell strain loading device, and the two dynamic positioning holes 2 are connected to the mover of the adherent cell strain loading device; during the loading process, the two static positioning holes 1 remain stationary, and the two dynamic positioning holes 2 are stretched with different displacements as they are driven; cell culture fluid is added to the upper end plane 3 of the elastic basement membrane to seed cells; the longitudinal cross-section of the lower end curved surface 4 of the elastic basement membrane is an "M"-shaped structure, and the distance from the upper end plane 3 of the elastic basement membrane is the thickness of the elastic basement membrane, and its thickness variation meets the uniformity requirement of the strain of the upper end plane 3 of the elastic basement membrane during the chamber loading process; During the stretching process, the thickness of the elastic basement membrane is continuously adjusted, and the relative position of the portion of the upper end plane 3 of the elastic basement membrane with greater strain, that is, the distance between the lower end curved surface 4 of the elastic basement membrane and the upper end plane 3 of the elastic basement membrane, that is, the thickness of the elastic basement membrane, is increased. During the actual loading process, the strain value of the upper end plane 3 of the elastic basement membrane will be reduced; for the portion of the upper end plane 3 of the elastic basement membrane with less strain, that is, the distance between the lower end curved surface 4 of the elastic basement membrane and the upper end plane 3 of the elastic basement membrane, that is, the thickness of the elastic basement membrane, will be increased during the actual loading process, thereby achieving the purpose of uniform strain on the upper end plane 3 of the elastic basement membrane.

2. The novel uniform strain chamber of the adherent cell loading device according to claim 1, characterized in that: The total horizontal length of the chamber is 70mm and the longitudinal length is 30mm. The longitudinal distance of the inner cavity is 26.8mm, the horizontal distance is 40mm, and the height is 15mm. The diameter of the two static positioning holes 1 is 4mm, and the diameter of the two dynamic positioning holes 2 is 4mm. The chamber has different structures under three different stretching rates of 1%, 5%, and 10%. The difference is the distance between the upper end plane 3 of the elastic basement membrane and the lower end curved surface 4 of the elastic basement membrane, that is, the thickness of the elastic basement membrane. The shape of any longitudinal cross-section of the lower end curved surface 4 of the elastic basement membrane is "M"-shaped.

3. The novel uniform strain chamber of the adherent cell loading device according to claim 2, characterized in that: When the stretching rate is 1%, the maximum thickness of the elastic basement membrane is 0.651 mm and the minimum thickness is 0.231 mm.

4. The novel uniform strain chamber of the adherent cell loading device according to claim 2, characterized in that: When the stretching rate is 5%, the maximum thickness of the elastic basement membrane is 0.692 mm and the minimum thickness is 0.314 mm.

5. The novel uniform strain chamber of the adherent cell loading device according to claim 2, characterized in that: When the stretching rate is 10%, the maximum thickness of the elastic basement membrane is 0.616 mm and the minimum thickness is 0.314 mm.

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