Bioactive glass particles as nucleic acid carriers for controlled release

Abstract

The invention relates to bioactive glass particles as nucleic acid carriers for controlled release. The glass powder includes a plurality of modified glass particles including, in weight percent (wt%): greater than or equal to 20 wt% to less than or equal to 75 wt% SiO2; from greater than or equal to 3 wt% to less than or equal to 25 wt% P2O5; caO greater than or equal to 2 wt% to less than or equal to 50 wt%; and the surface of the plurality of modified glass particles comprises a total amount of brushite and apatite greater than or equal to 30% by weight of the total weight of the modified glass particles, in which the plurality of modified glass particles bind to a nucleic acid compound after contact.

Description

Technical Field

[0001] The present disclosure relates to glass compositions for delivering nucleic acids into cells or microorganisms. In particular, the present disclosure relates to bioactive glass compositions capable of capturing and releasing nucleic acids in a specified environment. Background Art

[0002] The use or transfer of exogenous nucleic acids for gene regulation or control is an emerging market of interest, especially in the agricultural and therapeutic fields. The ability of small interfering RNA (siRNA) to regulate gene expression has potential therapeutic applications, but its use is limited by inefficient delivery. Specifically, due to the low stability and low cellular uptake efficiency of nucleic acids, the delivery of exogenous nucleic acids into target organisms or cells remains challenging. Therefore, there is a need for new glass compositions with improved surface properties that can act as nucleic acid carriers to enhance cellular uptake rates. Summary of the invention

[0003] The first aspect may relate to a glass powder comprising a plurality of modified glass particles, wherein the plurality of modified glass particles comprise, in weight percentage (wt%): greater than or equal to 20 wt% to less than or equal to 75 wt% SiO2, greater than or equal to 3 wt% to less than or equal to 25 wt% P2O5, greater than or equal to 2 wt% to less than or equal to 50 wt% CaO, and a total amount of brushite and apatite contained on the surfaces of the plurality of modified glass particles is greater than or equal to 30 wt% of the total weight of the modified glass particles, wherein the plurality of modified glass particles bind to a nucleic acid compound after contacting.

[0004] A second aspect of the present disclosure may include the first aspect, wherein the plurality of modified glass particles further comprises greater than 0 wt % to less than or equal to 5 wt % B2O3.

[0005] A third aspect of the present disclosure may include the first or second aspect, wherein the plurality of modified glass particles further comprises greater than 0 wt % to less than or equal to 5 wt % Al 2 O 3 .

[0006] A fourth aspect of the present disclosure may include any one of aspects 1 to 3, wherein the plurality of modified glass particles further comprises greater than 0 wt % to less than or equal to 30 wt % Na2O.

[0007] A fifth aspect of the present disclosure may include any one of aspects 1 to 4, wherein the plurality of modified glass particles further comprises greater than 0 wt % to less than or equal to 10 wt % K2O.

[0008] A sixth aspect of the present disclosure may include any one of aspects 1 to 5, wherein the plurality of modified glass particles further comprises greater than 0 wt % to less than or equal to 20 wt % MgO.

[0009] A seventh aspect of the present disclosure may include any one of aspects 1 to 6, wherein the plurality of modified glass particles further comprises greater than 0 wt % to less than or equal to 10 wt % Zr2O.

[0010] An eighth aspect of the present disclosure may include any one of aspects 1 to 7, wherein the plurality of modified glass particles further comprises greater than 0 wt % to less than or equal to 5 wt % F.

[0011] A ninth aspect of the present disclosure may include any one of aspects 1 to 8, wherein the surfaces of the plurality of modified glass particles include brushite in an amount greater than or equal to 10 weight % of the total weight of the modified glass particles, and include apatite in an amount greater than or equal to 15 weight % of the total weight of the modified glass particles.

[0012] A tenth aspect of the present disclosure may include any one of aspects 1 to 9, wherein the surfaces of the plurality of modified glass particles include brushite in an amount greater than or equal to 50 weight % of the total weight of the modified glass particles, and include apatite in an amount greater than or equal to 3 weight % of the total weight of the modified glass particles.

[0013] An 11th aspect of the present disclosure may include any one of aspects 1 to 10, wherein the total amount of brushite and apatite included on the surfaces of the plurality of modified glass particles is greater than or equal to 60 weight % of the total weight of the modified glass particles.

[0014] A twelfth aspect of the present disclosure may include any one of aspects 1 to 11, wherein the plurality of modified glass particles include greater than or equal to 100 m 2 / g to less than or equal to 300m 2 / g of BET surface area.

[0015] A 13th aspect of the present disclosure may include any one of aspects 1 to 12, wherein the plurality of modified glass particles include an average pore width of greater than or equal to 2 nanometers (nm) to less than or equal to 10 nm, as determined by BET.

[0016] A 14th aspect of the present disclosure may include any one of aspects 1 to 13, wherein the plurality of modified glass particles include a zeta potential of greater than or equal to -10 mV to less than or equal to 5 mV.

[0017] A 15th aspect of the present disclosure may include any one of the 1st to 14th aspects, wherein surfaces of the plurality of modified glass particles include needle-like crystals having a maximum length of greater than or equal to 10 nm to less than or equal to 50 nm.

[0018] A sixteenth aspect of the present disclosure may include any one of aspects 1 to 15, wherein the plurality of modified glass particles have an average longest dimension of greater than or equal to 0.5 micrometers (μm) to less than or equal to 10 μm.

[0019] A seventeenth aspect of the present disclosure may include any one of aspects one to sixteen, wherein the plurality of modified glass particles comprise: greater than or equal to 25 wt % to less than or equal to 50 wt % SiO2; greater than or equal to 8 wt % to less than or equal to 25 wt % P2O5; and greater than or equal to 30 wt % to less than or equal to 50 wt % CaO.

[0020] An eighteenth aspect of the present disclosure may include any one of aspects one to seventeen, wherein the plurality of modified glass particles comprise: greater than or equal to 25 weight percent to less than or equal to 35 weight percent SiO2; greater than or equal to 15 weight percent to less than or equal to 25 weight percent P2O5; greater than or equal to 3 weight percent to less than or equal to 6 weight percent MgO; greater than or equal to 38 weight percent to less than or equal to 48 weight percent CaO; and greater than or equal to 2 weight percent to less than or equal to 6 weight percent ZrO2.

[0021] A 19th aspect of the present disclosure may include any one of aspects 1 to 18, wherein the plurality of modified glass particles comprise: greater than or equal to 38 wt % to less than or equal to 48 wt % SiO2; greater than or equal to 5 wt % to less than or equal to 15 wt % P2O5; greater than or equal to 10 wt % to less than or equal to 20 wt % MgO; and greater than or equal to 28 wt % to less than or equal to 38 wt % CaO.

[0022] A 20th aspect of the present disclosure may include any one of aspects 1 to 19, wherein the plurality of modified glass particles further comprises a nucleic acid compound to obtain a bioactive glass carrier.

[0023] Aspect 21 of the present disclosure may include aspect 20 or any of the preceding aspects, wherein the nucleic acid compound comprises deoxyribonucleic acid (DNA) comprising less than or equal to 1500 base pairs, ribonucleic acid (RNA) comprising less than or equal to 1000 base pairs, or a combination thereof.

[0024] A 22nd aspect of the present disclosure may include any of the 20th or 21st aspects or any of the preceding aspects, wherein the concentration of the nucleic acid compound is greater than or equal to 500 nanograms per milligram (ng / mg) to less than or equal to 5,000 ng / mg, based on the total weight of the plurality of modified glass particles.

[0025] A 23rd aspect of the present disclosure may relate to a method for forming a glass powder, the method comprising: processing glass into precursor glass particles, wherein the precursor glass particles have an average longest dimension greater than or equal to 0.5 micrometers (μm) to less than or equal to 10 μm; and treating the precursor glass particles with a treatment solution, wherein: the treatment solution comprises a phosphate, and the treatment forms modified glass particles; wherein: the total amount of calcium apatite and apatite contained on the surface of the plurality of modified glass particles is greater than or equal to 30 weight % of the total weight of the modified glass particles, and the plurality of modified glass particles can be combined with a nucleic acid compound after contacting.

[0026] A 24th aspect of the present disclosure may include the 23rd aspect or any of the preceding aspects, wherein the pH of the treatment solution is greater than or equal to 2.5 to less than or equal to 4.5.

[0027] Aspect 25 of the present disclosure may include any one of aspect 23 or 24 or any of the preceding aspects, wherein: the processing includes forming a mixture comprising precursor glass particles and a processing solution; and the concentration of the phosphate in the mixture is greater than or equal to 0.1 molar (M) to less than or equal to 1M.

[0028] Aspect 26 of the present disclosure may include any one of aspects 23 to 25 or any of the preceding aspects, wherein the treatment solution comprises sodium phosphate, potassium phosphate, or a combination thereof.

[0029] A 27th aspect of the present disclosure may include any one of aspects 23 to 26 or any of the preceding aspects, wherein the processing includes: forming a mixture comprising precursor glass particles and a processing solution; and heating the mixture at a temperature of 40° C. to 80° C. for a heating duration.

[0030] Aspect 28 of the present disclosure may include aspect 27 or any preceding aspect, wherein the heating duration is greater than or equal to 1 day.

[0031] A 29th aspect of the present disclosure may relate to a method for forming a bioactive glass carrier, the method comprising: processing glass into precursor glass particles, wherein the precursor glass particles have an average longest dimension greater than or equal to 0.5 microns (μm) to less than or equal to 10 μm; and treating the precursor glass particles with a treatment solution, wherein: the treatment solution comprises a phosphate, and the treatment forms modified glass particles; contacting the plurality of modified glass particles with a nucleic acid compound to form a mixture; and incubating the mixture under conditions such that the nucleic acid compound non-covalently binds to the plurality of modified glass particles to form a bioactive glass carrier.

[0032] A 30th aspect of the present disclosure may include the 29th aspect or any preceding aspect, wherein the mixture comprises a nucleic acid compound at a concentration of greater than or equal to 500 ng per mg of the plurality of modified glass particles (ng / mg) to less than or equal to 10,000 ng / mg.

[0033] A 31st aspect of the present disclosure may include any one of the 29th or 30th aspects or any of the preceding aspects, wherein incubating the mixture is performed at a temperature of greater than or equal to 20°C to less than or equal to 40°C.

[0034] A 32nd aspect of the present disclosure may include any one of aspects 29 to 31 or any of the preceding aspects, wherein the mixture is incubated for a period of time greater than or equal to 10 minutes.

[0035] A 33rd aspect of the present disclosure may include any one of aspects 29 to 32 or any of the preceding aspects, wherein incubating the mixture includes loading the nucleic acid compound onto the plurality of modified glass particles at a loading efficiency greater than or equal to 50%.

[0036] Aspect 34 of the present disclosure may include any one of aspects 29 to 33 or any of the preceding aspects, wherein the bioactive glass carrier comprises a nucleic acid compound at a concentration of greater than or equal to 500 ng per mg of bioactive glass carrier (ng / mg) to less than or equal to 5,000 ng / mg.

[0037] A 35th aspect of the present disclosure may relate to a carrier comprising the glass powder (or bioactive glass carrier) according to any one of aspects 1 to 22 or any of the preceding aspects.

[0038] Aspect 36 of the present disclosure may include aspect 35 or any of the preceding aspects, wherein the carrier comprises: a drug delivery carrier, a gene delivery carrier, a tissue regeneration carrier, a crop treatment or crop protection carrier, an immunomodulatory carrier, or a gene editing complex carrier.

[0039] A 37th aspect of the present disclosure may include any of the 35th or 36th aspects or any of the preceding aspects, wherein at least a portion of the nucleic acid compound is released at a pH of greater than or equal to 3 to less than or equal to 10.

[0040] A 38th aspect of the present disclosure may include the 37th aspect or any of the preceding aspects, wherein at least 60 weight percent of the nucleic acid compound is released, based on the total weight of the nucleic acid compound loaded in the carrier.

[0041] Aspect 39 of the present disclosure may relate to a method for releasing nucleic acid compounds from a glass powder (or a bioactive glass carrier) of any one of aspects 20 to 22 or 35 to 38 or any of the preceding aspects, the method comprising: placing the glass powder (or the bioactive glass carrier) in a cellular environment, wherein at least a portion of the nucleic acid compound is released from the glass powder (or the bioactive glass carrier) in the cellular environment.

[0042] A 40th aspect of the present disclosure may include the 39th aspect or any preceding aspect, wherein at least a portion of the nucleic acid compound is released at a pH of greater than or equal to 3 to less than or equal to 10.

[0043] A 41st aspect of the present disclosure may include the 39th or 40th aspect or any of the preceding aspects, wherein at least 60 weight percent of the nucleic acid compound is released, based on the total weight of the nucleic acid compound in the glass powder (or bioactive glass carrier).

[0044] Aspect 42 of the present disclosure may include any one of aspects 39 to 41 or any of the preceding aspects, wherein the cellular environment comprises one or more of: insect cells, animal cells and embryos, fungi, fish, yeast, plant cells, or any combination thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 An X-ray diffraction (XRD) pattern of a modified glass particle according to one or more aspects shown and described herein is provided, as a function of intensity (y-axis) versus position (x-axis);

[0046] Figure 2A providing a scanning electron microscope (SEM) image of a precursor glass particle according to one or more aspects shown and described herein;

[0047] Figure 2B provides a scanning electron microscope (SEM) image of a modified glass particle according to one or more aspects shown and described herein;

[0048] Figure 2Cprovides a scanning electron microscope (SEM) image of a modified glass particle according to one or more aspects shown and described herein;

[0049] Figure 2D providing a scanning electron microscope (SEM) image of a precursor glass particle according to one or more aspects shown and described herein;

[0050] Figure 2E provides a scanning electron microscope (SEM) image of a modified glass particle according to one or more aspects shown and described herein;

[0051] Figure 2F provides a scanning electron microscope (SEM) image of a modified glass particle according to one or more aspects shown and described herein;

[0052] Figure 3 providing an electrophoretic image of a glass powder subjected to a DNase challenge according to one or more aspects shown and described herein; and

[0053] Figure 4 Provided are fluorescent and merged confocal images of glass powder internalized in cells according to one or more aspects shown and described herein. DETAILED DESCRIPTION

[0054] Specific reference will now be made to glass particles that can be used as nucleic acid carriers, various aspects of which are shown in the accompanying drawings. Whenever possible, the same reference numerals are used in the drawings to represent the same or similar parts. According to aspects, the glass powder can include a plurality of modified glass particles, which can be used as nucleic acid carriers. In terms of weight percentage (wt%), the plurality of modified glass particles can include: greater than or equal to 20 weight percent (wt%) to less than or equal to 75 wt% SiO2; greater than or equal to 3 wt% to less than or equal to 25 wt% P2O5; and greater than or equal to 2 wt% to less than or equal to 50 wt% CaO. The total amount of brushite and apatite contained on the surface of each of the plurality of modified glass particles can be greater than or equal to 30 wt% of the total weight of the modified glass particles, wherein the modified glass particles are combined with nucleic acid compounds after contact. Various aspects of glass particles used as nucleic acid carriers will be described herein with specific reference to the drawings. The components in the drawings are not necessarily to scale, but rather, are highlighted to show the principles of the exemplary aspects.

[0055] The present disclosure generally relates to glass powders, methods of forming glass powders, and methods of forming bioactive glass supports.

[0056] Definitions of certain terms used in this disclosure are provided below. Unless defined otherwise, all technical and scientific terms used herein generally have the meaning commonly understood by one of ordinary skill in the art to which this technology belongs.

[0057] As used in the present disclosure and claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a" component includes aspects having two or more such components unless the context clearly dictates otherwise. For example, reference to "a cell" includes combinations of two or more cells, and so forth. In addition, use of the word "or" when not preceded by the antecedent "either" (or when other similar terms indicating "or" are explicitly exclusive, e.g., only one of x or y) should be read as inclusive (e.g., "x or y" means one or both of x or y). In general, the nomenclature used herein and the cell culture, molecular genetics, organic chemistry, analytical chemistry, and nucleic acid chemistry described below are those well known and commonly used in the art.

[0058] The term "and / or" should also be interpreted as inclusive (e.g., "x and / or y" means one or both of x or y). Where "and / or" or "or" is used in relation to a group of three or more items, the group should be understood to include only one item, all items together, or any combination or number of items. In addition, terms such as: having, having, including, and including used in the specification and claims should be interpreted as having the same meaning as the terms including and comprising. With the "and / or" clause, other elements may optionally be present in addition to the specifically specified elements, whether or not they are related or unrelated to those specifically specified elements. As non-limiting examples, reference to "X and / or Y" may refer in one aspect to only X (optionally including elements other than Y); in some aspects to only Y (optionally including elements other than X); and in some aspects to both X and Y (optionally including other elements).

[0059] Unless otherwise indicated, as used herein, the term "particle size" refers to the longest dimension of a particle. "Average particle size" refers to the average of the longest dimensions of a plurality of particles. In aspects where the particles are spherical, the longest dimension is the particle diameter. For example, the particle size is expressed as the average particle size or the average longest dimension of a plurality of particles, which can be determined using electron microscopy in conjunction with image analysis.

[0060] As used herein, the term "bioactive glass" ("BAG") or "bioactive glass" refers to an inorganic glass material that is capable of binding to a desired biological material (e.g., nucleic acid). As used herein, the term "bioactive glass carrier" refers to a glass material that is bound to a desired biological material (e.g., nucleic acid).

[0061] As used herein, the term "controlled release" refers to the release of nucleic acids from a bioactive glass carrier according to a predetermined profile. In some aspects, the selected controlled release may include the release of nucleic acid compounds within a specific pH range, ionic strength range (or both).

[0062] Various aspects and / or embodiments of the present disclosure relate to glass powders comprising modified glass particles. In various aspects, the total amount of brushite and apatite contained on the surface of the modified glass particles is greater than or equal to 30% by weight of the total weight of the modified glass particles. The modified glass particles are capable of binding with nucleic acids after contact. The glass powder disclosed herein exhibits excellent biocompatibility. The bioactive properties of the glass powder are derived from a series of reactions on the surface of the precursor glass particles under certain conditions. For example, when the precursor glass particles are exposed to a treatment solution containing phosphate, cation exchange may occur, whereby cations from the glass surface are replaced by cations from the treatment solution. Ca released from the glass surface due to the dissolution of the glass network 2+ Ion-bound PO4 in solution 3- The ions react on the glass surface. This promotes the formation of brushite (CaHPO4·2H2O), a known bioactive ceramic, or apatite (e.g., hydroxyapatite (Ca 10 (PO4)6(OH)2)). Both brushite and apatite are capable of binding to nucleic acids (e.g., DNA and RNA). In aspects, apatite may refer to hydroxyapatite, fluorapatite (Ca5(PO4)3F), chloroapatite (Ca5(PO4)3F), or a combination thereof. In aspects, apatite may refer to hydroxyapatite.

[0063] A glass powder comprising a plurality of modified glass particles may be formed. In aspects, modified glass particles of the glass powder are formed from precursor glass particles. The glass powder may exhibit excellent nucleic acid capture / release efficiency.

[0064] The formation method of glass powder may include processing glass into precursor glass particles. Depending on the composition, the glass can be melted in a temperature range of about 1000°C to about 1600°C. For example, the glass can be melted in a temperature range of about 1100°C to about 1500°C or even in a temperature range of about 1150°C to about 1450°C. The glass can be melted in a suitable device (e.g., a covered platinum crucible) for a sufficient time to melt the glass, such as about 60 minutes (min) to about 24 hours (h), including: about 90min to about 20h, about 2h to about 18h, about 5h to about 16h, about 8h to about 12h, or any range between any two of these values ​​and / or any two of these values. After melting and clarification, the glass can be poured and annealed at a temperature of about 350°C to about 550°C (e.g., about 400°C to about 500°C). Then, the annealed glass can be poured into a suitable mold, and the resulting material is then cooled to form glass. In certain aspects, the glass can be ground into precursor glass particles having a particle size of 0.5 to 10 μm by air jet milling. Using friction milling or ball milling, the particle size will vary in the range of 1-100 μm. In addition, these glasses can be subsequently processed to submicron particle sizes using different methods (e.g., planetary ball milling or wet milling). In aspects, the precursor glass particles have an average longest dimension (particle size) greater than or equal to 0.5 microns (μm) to less than or equal to 10.0 μm, for example: 0.5 μm to 9.0 μm, 0.6 μm to 8.0 μm, 0.7 μm to 7.0 μm, 0.8 μm to 6.0 μm, 1.0 μm to 5.0 μm, 2.0 μm to 4.0 μm, or any range including any two of these values ​​and / or any range between any two of these values. For industrial scale, a top-down melting scheme can be used after grinding or milling to produce precursor glass particles of desired size, which provides flexibility in composition design and low-cost manufacturing. In certain embodiments, the glass can be ground into precursor glass particles using air jets, ball milling, or grinding. The precursor glass particles of desired size can be further processed to form modified glass particles.

[0065] The method of forming a glass powder may include treating precursor glass particles with a treatment solution, wherein the treatment solution comprises a phosphate, thereby forming modified glass particles. In one aspect, the treatment solution comprises a phosphate, such as but not limited to sodium sulfate, potassium sulfate, or a combination thereof. In one aspect, the treatment solution is an aqueous solution.

[0066] In aspects, the pH of the treatment solution can be 2.5 to 5.0, for example: 2.5 to 4.8, 3.0 to 4.5, 3.5 to 4.5, or any range comprising any two of these values ​​and / or any range between any two of these values. Without intending to be bound by any particular theory, it is believed that treating the precursor glass particles with a treatment solution having a pH less than 2.5 can cause at least a portion of the precursor glass particles to dissolve. In addition, it is believed that treating the precursor glass particles with a treatment solution having a pH greater than 5.0 can reduce the formation of apatite.

[0067] The treatment can include forming a mixture comprising the precursor glass particles and the treatment solution. In one aspect, the concentration of the phosphate in the mixture can be 0.1 molar (M) to 1 M, for example: 0.2 M to 0.8 M, 0.3 M to 0.7 M, 0.4 M to 0.6 M, or any range including any two of these values ​​and / or any range between any two of these values.

[0068] In aspects, the treatment can include heating the mixture at an elevated temperature (e.g., 40° C. to 80° C. or 50° C. to 70° C.) for a heating duration. In aspects, the heating duration is greater than or equal to 1 day, greater than or equal to 3 days, or greater than or equal to 5 days. In aspects, the heating duration is 1 day to 60 days, 1 day to 30 days, 1 day to 15 days, 1 day to 7 days, 3 days to 60 days, 3 days to 30 days, 3 days to 15 days, 3 days to 10 days, or 3 days to 7 days. Without intending to be bound by any particular theory, it is believed that heating durations shorter than 1 day may not provide for sufficient formation of apatite and brushite on the surfaces of the modified glass particles.

[0069] Treating the precursor glass particles with a treatment solution can form modified glass particles having an increased concentration of brushite and apatite on the surface of the modified glass particles (this is relative to the concentration of brushite and apatite on the surface of the precursor glass particles). In addition, the treatment can increase the surface area of ​​the modified glass particles. The increase in both the surface area of ​​the modified glass particles and the concentration of brushite and apatite can increase the nucleic acid loading capacity of the modified glass particles.

[0070] In one aspect, a method for forming a bioactive glass carrier may include: contacting modified glass particles with a nucleic acid compound to form a mixture, and incubating the mixture under conditions such that the nucleic acid compound is non-covalently bound to the modified glass particles to form the bioactive glass carrier. In one aspect, the mixture may include a suitable liquid medium, such as water, alcohols, or hydrocarbons.

[0071] The concentration of the nucleic acid compound included in the mixture can be 500 nanograms (ng) per milligram (mg) of modified glass particles (ng / mg) to 10,000 ng / mg. For example, the concentration of the nucleic acid compound included in the mixture can be: 500 ng / mg to 10,000 ng / mg, 750 ng / mg to 8,000 ng / mg, 1,000 ng / mg to 6,000 ng / mg, 1,250 ng / mg to 4,000 ng / mg, 1,500 ng / mg to 3,000 ng / mg, or any range including any two of these values ​​and / or any range between any two of these values.

[0072] In aspects, incubating the mixture can include treating the mixture at a temperature of 20°C to 40°C (e.g., 20°C to 30°C or 20°C to 25°C). Without intending to be limited to any particular theory, it is believed that incubating the mixture at a temperature greater than 40°C may destabilize the nucleic acid. Incubation can be performed for a period of time greater than or equal to 10 minutes. In aspects, incubation can be performed for a period of time less than or equal to 3 hours, less than or equal to 2 hours, or less than or equal to 1 hour.

[0073] The loading efficiency of the nucleic acid loaded on the modified glass particles can be determined as follows: the amount of nucleic acid in the mixture (adding total nucleic acid) is calculated, and the amount of residual nucleic acid in the supernatant of the mixture after the modified glass particles (unloaded nucleic acid) is calculated. The amount of nucleic acid loaded can be determined by subtracting the amount of residual nucleic acid in the supernatant from the total amount of nucleic acid in the mixture. The loading efficiency is calculated by dividing the amount of nucleic acid loaded by the total amount of nucleic acid in the mixture. The concentration of nucleic acid in the mixture and the supernatant can be determined using conventional methods in the art (e.g., using a microvolume UV-visible spectrophotometer or a fluorescent Qubit nucleic acid assay). The surface charge and capacity of the modified glass particles may affect the loading efficiency. In aspect, incubating the mixture includes loading the nucleic acid compound onto the modified glass particles with a loading efficiency greater than or equal to 50%, for example: greater than or equal to 70%, greater than or equal to 80%, or even greater than or equal to 90%. In aspect, the loading efficiency can be 50% to 100%, for example: 60% to 95%, 70% to 95%, 80% to 95%, or 90% to 95%.

[0074] As discussed herein, a glass powder can include a plurality of modified glass particles. In aspects, the modified glass particles of the glass powder are formed from precursor glass particles. The biocompatibility and stability of the glass powder are affected by the composition of the modified glass particles.

[0075] In one aspect, the precursor glass particles or the modified glass particles of the glass powder formed therefrom contain SiO2. When the content of SiO2 is too low, the glass may have poor chemical durability and mechanical durability. On the other hand, when the content of SiO2 is too high, the melting ability of the glass may decrease because the melting temperature of pure SiO2 is high. As the content of SiO2 increases, the biological activity of the modified glass particles may decrease. In one aspect, the precursor glass particles or the modified glass particles of the glass powder formed therefrom include silicate glass, which contains SiO2 as the main glass-forming oxide. In one aspect, the amount of SiO2 present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is 20 wt% to 75 wt%, for example: 25 wt% to 70 wt%, 30 wt% to 65 wt%, 35 wt% to 60 wt%, 40 wt% to 55 wt%, 45 wt% to 50 wt%, respectively, relative to the total weight of the precursor glass particles and the modified glass particles, or any range including any two of these values ​​and / or between any two of these values, based on the total weight of the precursor glass particles or the modified glass particles.

[0076] The precursor glass particles or the modified glass particles of the glass powder formed therefrom may also contain P2O5, which acts as a network former. In addition, the release of phosphate ions to the surface of the precursor glass particles may contribute to the formation of calcium-phosphorus (Ca-P) compounds (e.g., brushite or apatite). In addition, P2O5 may increase the viscosity of the glass, which in turn expands the operating temperature range and may be advantageous for the manufacture and formation of glass subsequently processed into glass powder. In various aspects, the amount of P2O5 present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is 3% to 25% by weight of the total weight of the precursor glass particles or the modified glass particles, for example: 8% to 25% by weight, 15% to 25% by weight, or 5% to 15% by weight of P2O5. In one aspect, the precursor glass particles or the modified glass particles of the glass powder formed therefrom contain PO in an amount of 3 wt % to 25 wt %, 5 wt % to 20 wt %, 8 wt % to 15 wt %, 10 wt % to 12 wt %, or any range including any two of these values ​​and / or any range between any two of these values, based on the total weight of the precursor glass particles or the modified glass particles.

[0077] In one aspect, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may contain divalent cation oxides (e.g., alkaline earth oxides), which improve the melting behavior of the glass. The divalent cations may also improve the bioactivity of the glass. CaO may react with P2O5 in the precursor glass particles to form apatite. In addition, CaO may react with a treatment solution containing phosphate to form apatite. The CaO released from the surface of the precursor glass particles may be used to form apatite. 2+ The ions can contribute to the formation of a surface layer of the particles enriched with calcium phosphate. In aspects, the amount of CaO present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is 2 wt % to 50 wt %, based on the total weight of the precursor glass particles or the modified glass particles. For example, in aspects, the amount of CaO present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom includes: 30 wt % to 50 wt %, 28 wt % to 38 wt %, or 38 wt % to 48 wt % CaO, based on the total weight of the precursor glass particles or the modified glass particles. In aspects, the amount of CaO present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is: 2 wt % to 50 wt %, 5 wt % to 45 wt %, 10 wt % to 40 wt %, 15 wt % to 35 wt %, 20 wt % to 30 wt %, 28 wt % to 38 wt %, 38 wt % to 48 wt %, or any range including any two of these values ​​and / or between any two of these values, based on the total weight of the precursor glass particles or the modified glass particles.

[0078] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may contain B2O3, which may increase the biological activity of the modified glass particles of the glass powder. In aspects, the amount of B2O3 present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is 0 wt % to 5 wt %, based on the total weight of the precursor glass particles or the modified glass particles. For example, in aspects, the amount of B2O3 present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom includes: 0 wt % to 5 wt %, 0.2 wt % to 4.5 wt %, 0.5 wt % to 4 wt %, 1 wt % to 3.5 wt %, 1.5 wt % to 3 wt %, 2 wt % to 2.5 wt %, or includes any two of these values ​​and / or any range between any two of these values, based on the total weight of the precursor glass particles or the modified glass particles.

[0079] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may contain Al2O3, which may increase the thermal stability of the glass. In aspects, the amount of Al2O3 present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is 0 wt% to 5 wt%, based on the total weight of the precursor glass particles or the modified glass particles. For example, in aspects, the amount of Al2O3 present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom includes: 0 wt% to 5 wt%, 0.2 wt% to 4.5 wt%, 0.5 wt% to 4 wt%, 1 wt% to 3.5 wt%, 1.5 wt% to 3 wt%, 2 wt% to 2.5 wt%, or includes any two of these values ​​and / or any range between any two of these values, based on the total weight of the precursor glass particles or the modified glass particles.

[0080] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may contain Na2O, which may increase the bioactivity of the glass. In aspects, the amount of Na2O present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is 0 wt % to 30 wt %, based on the total weight of the precursor glass particles or the modified glass particles. For example, in aspects, the amount of Na2O present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom includes: 0 wt % to 30 wt %, 5 wt % to 25 wt %, 10 wt % to 20 wt %, 12 wt % to 15 wt %, or includes any two of these values ​​and / or any range between any two of these values, based on the total weight of the precursor glass particles or the modified glass particles.

[0081] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may contain K2O, which may increase the bioactivity of the glass. In aspects, the amount of K2O present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is 0 wt % to 10 wt %, based on the total weight of the precursor glass particles or the modified glass particles. For example, in aspects, the amount of K2O present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom includes: 0 wt % to 10 wt %, 1 wt % to 9 wt %, 2 wt % to 8 wt %, 3 wt % to 7 wt %, 4 wt % to 6 wt %, 4 wt % to 5 wt %, or includes any two of these values ​​and / or any range between any two of these values, based on the total weight of the precursor glass particles or the modified glass particles.

[0082] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may contain MgO, which may increase the bioactivity of the glass. In aspects, the amount of MgO present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is 0 wt % to 20 wt %, based on the total weight of the precursor glass particles or the modified glass particles. For example, in aspects, the amount of MgO present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom includes: 0 wt % to 20 wt %, 1 wt % to 18 wt %, 2 wt % to 16 wt %, 3 wt % to 15 wt %, 5 wt % to 15 wt %, 6 wt % to 14 wt %, 7 wt % to 13 wt %, 8 wt % to 12 wt %, 9 wt % to 11 wt %, 10 wt % to 20 wt %, 3 wt % to 6 wt %, or any two of these values ​​and / or any range between any two of these values, based on the total weight of the precursor glass particles or the modified glass particles.

[0083] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may contain ZrO2, which can increase the thermal stability of the glass. In aspects, the amount of ZrO2 present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is 0 wt % to 10 wt %, based on the total weight of the precursor glass particles or the modified glass particles. For example, in aspects, the amount of ZrO2 present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom includes: 0 wt % to 10 wt %, 1 wt % to 9 wt %, 2 wt % to 8 wt %, 3 wt % to 7 wt %, 4 wt % to 6 wt %, 4 wt % to 5 wt %, 2 wt % to 6 wt %, or includes any two of these values ​​and / or any range between any two of these values, based on the total weight of the precursor glass particles or the modified glass particles.

[0084] In one aspect, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may contain fluorine (F - ), which can increase the bioactivity of the glass. In one aspect, the F present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom - The amount of F present in the precursor glass particles or the modified glass particles is 0 wt % to 5 wt %, based on the total weight of the precursor glass particles or the modified glass particles. For example, in the aspect, the F present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom is -The amount includes: 0 wt % to 5 wt %, 0.2 wt % to 4.5 wt %, 0.5 wt % to 4 wt %, 1 wt % to 3.5 wt %, 1.5 wt % to 3 wt %, about 2 wt % to about 2.5 wt %, or any range including any two of these values ​​and / or any range between any two of these values, based on the total weight of the precursor glass particles or the modified glass particles.

[0085] In one aspect, the modified glass particles include: greater than or equal to 25 wt % to less than or equal to 50 wt % SiO2; greater than or equal to 8 wt % to less than or equal to 25 wt % P2O5; and greater than or equal to 30 wt % to less than or equal to 50 wt % CaO.

[0086] In one aspect, the modified glass particles include: greater than or equal to 25 wt% to less than or equal to 35 wt% SiO2; greater than or equal to 15 wt% to less than or equal to 25 wt% P2O5; greater than or equal to 3 wt% to less than or equal to 6 wt% MgO; greater than or equal to 38 wt% to less than or equal to 48 wt% CaO; and greater than or equal to 2 wt% to less than or equal to 6 wt% ZrO2.

[0087] In one aspect, the modified glass particles include: greater than or equal to 38 wt% to less than or equal to 48 wt% SiO2; greater than or equal to 5 wt% to less than or equal to 15 wt% P2O5; greater than or equal to 10 wt% to less than or equal to 20 wt% MgO; and greater than or equal to 28 wt% to less than or equal to 38 wt% CaO.

[0088] Modified glass particles formed from precursor glass particles may exhibit certain properties that are different from the precursor glass particles, such as, but not limited to, brushite and apatite surface concentrations, BET surface area, average pore width, or surface charge.

[0089] The concentration of brushite and / or apatite on the surface of the modified glass particles can be determined by quantitative phase analysis by XRD using the Rietveld method known in the art.

[0090] In one aspect, the amount of brushite included on the surface of the modified glass particles can be greater than or equal to 5% by weight of the total weight of the modified glass particles. For example, the amount of brushite included on the surface of the modified glass particles can be greater than or equal to 10% by weight, greater than or equal to 20% by weight, greater than or equal to 30% by weight, greater than or equal to 40% by weight, greater than or equal to 50% by weight, greater than or equal to 60% by weight, or greater than or equal to 70% by weight of the total weight of the modified glass particles. The amount of brushite included in the modified glass particles can be 5% to 90% by weight, 10% to 80% by weight, 20% to 70% by weight, 30% to 50% by weight of the total weight of the modified glass particles, or any range including any two of these values ​​and / or any range between any two of these values.

[0091] In one aspect, the amount of apatite included on the surface of the modified glass particles can be greater than or equal to 2 weight percent of the total weight of the modified glass particles. For example, the amount of apatite included on the surface of the modified glass particles can be greater than or equal to 3 weight percent, greater than or equal to 4 weight percent, greater than or equal to 5 weight percent, greater than or equal to 8 weight percent, or greater than or equal to 10 weight percent of the total weight of the modified glass particles. The amount of apatite included in the modified glass particles can be 2 weight percent to 40 weight percent, 3 weight percent to 30 weight percent, 4 weight percent to 20 weight percent, 5 weight percent to 15 weight percent of the total weight of the modified glass particles, or any range including any two of these values ​​and / or any range between any two of these values.

[0092] In one aspect, the total amount of brushite and apatite included on the surface of the modified glass particles can be greater than or equal to 30% by weight of the total weight of the modified glass particles. For example, the total amount of brushite and apatite included on the surface of the modified glass particles can be greater than or equal to 35% by weight, greater than or equal to 40% by weight, greater than or equal to 50% by weight, greater than or equal to 60% by weight, greater than or equal to 70% by weight, or even greater than or equal to 80% by weight of the total weight of the modified glass particles. The total amount of brushite and apatite included in the modified glass particles can be 30% to 95% by weight, 35% to 90% by weight, 40% to 85% by weight, 45% to 80% by weight, 50% to 75% by weight, 55% to 70% by weight, 60% to 65% by weight of the total weight of the modified glass particles, or any range including any two of these values ​​and / or any range between any two of these values.

[0093] In certain aspects, the surface of the modified glass particles may include brushite in an amount greater than or equal to 10% by weight of the total weight of the modified glass particles, and the amount of apatite may be greater than or equal to 15% by weight of the total weight of the modified glass particles. In other aspects, the surface of the modified glass particles may include brushite in an amount greater than or equal to 50% by weight of the total weight of the modified glass particles, and the amount of apatite may be greater than or equal to 3% by weight of the total weight of the modified glass particles.

[0094] In aspects, the weight percentage of apatite disclosed herein may refer to the weight percentage of hydroxyapatite.It should be understood that all ranges and subranges referring to the amount of apatite on the surface of the modified glass particles also disclose the amount of hydroxyapatite on the surface of the modified glass particles.

[0095] Without intending to be bound by any particular theory, it is believed that modified glass particles having increased amounts of brushite and / or apatite on the surface of the modified glass particles can increase the nucleic acid loading capacity of the modified glass particles.

[0096] Modified glass particles may include 100m 2 / g to 300m 2 / g BET surface area, for example: 100m 2 / g to 250m 2 / g,100m 2 / g to 200m 2 / g,125m 2 / g to 300m 2 / g,125m 2 / g to 250m 2 / g,125m 2 / g to 200m 2 / g,150m 2 / g to 300m 2 / g,150m 2 / g to 250m 2 / g,150m 2 / g to 200m 2 / g, or any range comprising any two of these values ​​and / or any two of these values. As used herein, "BET surface area" refers to the surface area determined by the Brunauer-Emmett-Teller theory. The surface area of ​​the modified glass particles can also be changed by changing the particle size. Without intending to be limited to any particular theory, it is believed that the modified glass particles with increased BET surface area can increase the nucleic acid loading capacity of the modified glass particles.

[0097] The modified glass particles can include an average pore width of 2 nm to 10 nm, for example, 3 nm to 9 nm, 4 nm to 8 nm, 5 nm to 7 nm, or any range including any two of these values ​​and / or between any two of these values ​​(determined by BET).

[0098] The modified glass particles may include a zeta potential of greater than or equal to -10 mV to less than or equal to 5 mV, for example: -8 mV to 4 mV, -5 mV to 4 mV, -2 mV to 3 mV, or any range comprising any two of these values ​​and / or any range between any two of these values. Without intending to be limited to any particular theory, it is believed that a surface charge close to neutral (neutral is 0 mV zeta potential) such as the zeta potential range described herein demonstrates lower active oxide exposure of the modified glass particles. Zeta potential can be measured by conventional means in the art (e.g., electrophoretic light scattering (Malvern Zetasizer Nano ZS)).

[0099] In aspects, the modified glass particles can include needle-like crystals of apatite and / or brushite with a maximum length greater than or equal to 10 nm to less than or equal to 50 nm. Without intending to be limited to any particular theory, it is believed that the needle-like crystals on the surface of the modified glass particles can provide nanometer-sized pores for nucleic acid binding, which can increase the nucleic acid loading capacity of the modified glass particles. The presence of needle-like crystals and the maximum length of the needle-like crystals can be determined by conventional means in the art (e.g., electron microscopy).

[0100] In aspects, the modified glass particles can have an average longest dimension of greater than or equal to 0.5 micrometers (μm) to less than or equal to 10.0 μm, for example: 0.5 μm to 9.0 μm, 0.6 μm to 8.0 μm, 0.7 μm to 7.0 μm, 0.8 μm to 6.0 μm, 1.0 μm to 5.0 μm, 2.0 μm to 4.0 μm, or any range including any two of these values ​​and / or any range between any two of these values. The average longest dimension of the modified glass particles can be determined by conventional means in the art (e.g., electron microscopy combined with image analysis).

[0101] As described herein, the modified glass particles can be combined with nucleic acid compounds after contact. In aspects, the modified glass particles can contain nucleic acid compounds. The nucleic acid compounds can include DNA containing less than or equal to 1500 base pairs, RNA containing less than or equal to 1000 base pairs, or a combination thereof. The modified glass particles containing nucleic acid compounds can be referred to as bioactive glass carriers.

[0102] In aspects, the bioactive glass carrier can contain a nucleic acid compound at a concentration of 500 ng per mg of bioactive glass carrier (ng / mg) to 5,000 ng / mg. For example, the bioactive glass carrier can contain a nucleic acid compound at a concentration of 500 ng / mg to 5,000 ng / mg, 750 ng / mg to 3,000 ng / mg, 1,000 ng / mg to 2,500 ng / mg, 1,500 ng / mg to 2,000 ng / mg, or any range including any two of these values ​​and / or any range between any two of these values.

[0103] Modified glass particles can be combined with nucleic acid compounds under various pH conditions. Nucleic acid mixtures can be coated on or located on the surface of bioactive glass particles. Suitable nucleic acids include any ribonucleic acid (RNA) or deoxyribonucleic acid (DNA), which can be modified or unmodified. In various aspects, nucleic acid compounds include, but are not limited to: single-stranded or double-stranded RNA, single-stranded or double-stranded DNA, RNA of a mixture of single-stranded and double-stranded regions, DNA of a mixture of single-stranded and double-stranded regions, mixed molecules containing DNA and RNA (can be single-stranded, or double-stranded, or a mixture of single-stranded and double-stranded regions). In addition, nucleic acid mixtures can refer to triple-stranded regions containing RNA or DNA or both of them, DNA or RNA containing one or more modified bases, and DNA or RNA with modified main chains for stability or other reasons. In some aspects, nucleic acid compounds include ribonucleic acid, deoxyribonucleic acid, or derivatives thereof. In some aspects, nucleic acid compounds include ribonucleic acid or derivatives thereof. In other aspects, nucleic acid compounds include deoxyribonucleic acid or derivatives thereof. In some aspects, nucleic acid compounds include interfering ribonucleic acid molecules.

[0104] In addition to apatite and brushite concentrations, some properties that verify the ability and capacity of nucleic acids loaded on modified glass particles can be surface charge and surface area. Surface charge can be directly determined by zeta potential measurement. The more positive charges on the particle surface, the higher the potential for capturing negative charges of nucleic acid compounds. On the other hand, the capture and release of nucleic acid compounds can be controllable and reversible. The carried nucleic acid can be released while being delivered to the target. It is known that the pH value inside the cell is slightly acidic compared to body fluids. Therefore, the surface negative charge of the carrier in the acidic solution can promote nucleic acid release. The nucleic acids bound on the surface of the modified glass particles can be evaluated by the change of the zeta potential of the modified glass particle surface. The change of zeta potential can indicate the ligand binding to the surface of the modified glass particles. Zeta potential is a surface charge measurement on modified glass particles and may have an impact on the stability of particles under various pH conditions.

[0105] Another aspect of the present disclosure relates to a method for releasing a nucleic acid compound from a glass powder (e.g., an active glass carrier) or a carrier comprising a glass powder (or an active glass carrier) as described herein. In aspects, the method may include placing the glass powder (or a bioactive glass carrier) in a cellular environment, wherein at least a portion of the nucleic acid compound is released from the glass powder (or a bioactive glass carrier) in the cellular environment. In aspects, the cellular environment may have a pH of 3 to 10, for example: 4 to 9, 5 to 8, 6 to 7, or any range comprising any two of these values ​​and / or any two of these values, or any other pH value or condition disclosed herein.

[0106] In aspects, the nucleic acid compound can be released at a rate of 60% by weight or more, for example: 65% by weight or more, 70% by weight or more, 75% by weight or more, or 80% by weight or more, based on the total weight of the nucleic acid compound loaded in the glass powder (or bioactive glass carrier). The nucleic acid compound released in the cell environment can be delivered to the target cell in the cell environment. It is not intended to be limited to any particular theory, but it is believed that the controlled release of nucleic acid compounds from glass powder (or bioactive glass carrier) in the cell environment can selectively increase the concentration of nucleic acid compounds in the target cell or near the target cell in the cell environment, thereby increasing the efficiency of delivering nucleic acid compounds to the desired cell environment. In some aspects, the cell environment includes one or more of: insect cells, animal cells and embryos, fungi, fish, yeast, plant cells, or any combination thereof.

[0107] Another aspect of the present disclosure relates to a carrier comprising a glass powder comprising modified glass particles described herein. The carrier may include: a drug delivery carrier, a gene delivery carrier, a tissue regeneration carrier, a crop treatment or crop protection carrier, an immunomodulatory carrier, or a gene editing composite carrier. The carrier can be used to deliver the modified glass particles and / or bioactive glass carriers described herein to various types of cells. According to the present disclosure, any type of cell type that is easy to transfer nucleic acids can be used as a target for transmission. Examples include, but are not limited to: insect cells, animal cells and embryos, fungi, fish, yeast, and plant cells. Stable aqueous suspensions can be used as part of a plant quarantine composition for controlling pests. For example, a stable suspension can be used to protect crops from insects, in which case the nucleic acid can be an insecticidal double-stranded RNA. Using interfering ribonucleic acid (RNA) molecules, the carrier can be used to prevent and / or control insect pest infestations in plants. Compositions and combinations containing interfering RNA molecules of the present technology can be used for topical applications, such as in the form of insecticides.

[0108] In aspects, at least a portion of the nucleic acid compound of the carrier can be released in an environment with a pH of 3 to 10, for example: 4 to 9, 5 to 8, 6 to 7, or any range comprising any two of these values ​​and / or any range between any two of these values. In aspects, greater than or equal to 60% by weight of the nucleic acid compound can be released, for example: greater than or equal to 65% by weight, greater than or equal to 70% by weight, greater than or equal to 75% by weight, or greater than or equal to 80% by weight, based on the total weight of the nucleic acid compound loaded in the carrier.

[0109] It will be apparent to those skilled in the art that various modifications and variations may be made without departing from the spirit or scope of the claimed subject matter. Therefore, the claimed subject matter is not to be restricted except by the appended claims and their equivalents.

[0110] Example

[0111] Various aspects will be further illustrated by the following examples, which are not intended to limit the present disclosure in any way.

[0112] Example 1: Preparation of initial glass powder

[0113] The exemplary glasses described herein are silicate glasses including a base composition comprising the constituent components listed in Table 1, in terms of weight percent of oxides.

[0114] Table 1

[0115] Oxide (weight %) Example 1-1 Example 1-2 <![CDATA[SiO2]]> 29.5 42.8 <![CDATA[B2O3]]> 0 0 <![CDATA[P2O5]]> 19.5 9.5 <![CDATA[Al2O3]]> 0 0 <![CDATA[Na2O]]> 0 0 <![CDATA[K2O]]> 0 0 MgO 4.5 14.4 CaO 42.5 32.4 <![CDATA[ZrO2]]> 4.0 0 <![CDATA[F - ]]> 0 0.8 Annealing point(℃) 650 400

[0116] Glass was made in a platinum crucible using the raw material batches listed in Table 1. Each crucible containing the raw material batches was placed in a preheated furnace and melted at 1100-1500° C. for 16 hours. The glass was refined to produce a molten precursor glass, which was then cast into a 7 inch by 11 inch by 0.5 inch block. The precursor glass was annealed at a temperature range of 400-650° C., as recorded in Table 1. The glass was crushed using air jets and ball mills to form initial glass powders (precursor glass particles), designated as Example 1-1 and Example 1-2.

[0117] Example 2: Acid Phosphate Treatment of Initial Glass Powder

[0118] The initial glass powders of Example 1-1 and Example 1-2 were modified by treating the initial glass powders in a 0.5M acidic phosphate solution in water (60°C) to form glass powders containing modified glass particles. During the treatment, the initial glass powders used and the acidic phosphate solution of each example of Example 2 are shown in Table 2.

[0119] Table 2

[0120] Example Initial glass powder Acid phosphate solution Processing time (days) Example 2-1 Example 1-1 <![CDATA[KH2PO4]]> 3 Example 2-2 Example 1-1 <![CDATA[NaH2PO4]]> 3 Example 2-3 Example 1-2 <![CDATA[KH2PO4]]> 7 Embodiment 2-4 Example 1-2 <![CDATA[NaH2PO4]]> 7

[0121] The crystal phases of the crystal phase assemblies and / or the crystal size of the crystal phases of the glass powders of Examples (Ex) 2-1 to 2-4 were determined by X-ray diffraction (XRD) analysis techniques known to those skilled in the art, using such commercially available instruments as a Bruker D4 Endeavor diffractometer manufactured by Bruker Corporation, Massachusetts, United States. The spectra were typically obtained from 10 to 100 degrees of 2θ. The glass powders of Examples 2-1 to 2-4 were subjected to XRD analysis, and Table 2 shows the results obtained. Figure 1 The XRD patterns of Example 2-1 (110), Example 2-2 (120), Example 2-3 (130), and Example 2-4 (140) are shown. Brushite (CaHPO4·2H2O) (shown as a peak marked with a circle) and hydroxyapatite (Ca 10 (PO4)6(OH)2) (shown as peaks marked with triangles) indicates the presence of both brushite and apatite on the surface of the modified glass particles, indicating the good biocompatibility of these modified glass particles. Quantitative phase analysis of the XRD patterns using the Rietveld method was used to determine the weight % of brushite and apatite at the surface of the modified glass particles relative to the total weight of the modified glass particles, and is summarized in Table 3.

[0122] Table 3

[0123]

[0124] The particle size and morphology of the glass compositions of Example 1 and Example 2 were analyzed by scanning electron microscopy (SEM, JEOL JSM-6500F with EDS detector). The SEM images are shown in FIG2 . Figure 2A Example 1-1 (untreated) is shown, Figure 2B Display Example 2-1, Figure 2C Display Example 2-2, Figure 2D Showing Example 1-2 (untreated), Figure 2E Showing Examples 2-3, and Figure 2F Examples 2-4 are shown. The average particle size of the examples is summarized in Table 4.

[0125] Table 4

[0126] Example Initial glass powder Acid phosphate solution Average particle size (μm) Example 1-1 Example 1-1 Untreated 2.3±0.9 Example 2-1 Example 1-1 <![CDATA[KH2PO4]]> 2.0±0.6 Example 2-2 Example 1-1 <![CDATA[NaH2PO4]]> 5.9±3.1 Example 1-2 Example 1-2 Untreated 1.43±0.42 Example 2-3 Example 1-2 <![CDATA[KH2PO4]]> 4.1±1.2 Embodiment 2-4 Example 1-2 <![CDATA[NaH2PO4]]> 1.1±0.5

[0127] The surface charge of the initial glass powder of Example 1 and the glass powder containing the modified glass particles of Example 2 were examined by measuring the zeta potential of the samples (Malvern Zetasizer Nano ZS). The samples for measuring the zeta potential were prepared by suspending the glass particles in ethanol. The zeta potentials of the initial glass powder and the acidic phosphate treated glass powders are shown in Table 5.

[0128] Table 5

[0129] Example Initial glass powder Acid phosphate solution Zeta potential(mV) Example 1-1 Example 1-1 Untreated 0.4 Example 2-1 Example 1-1 <![CDATA[KH2PO4]]> -7.2 Example 2-2 Example 1-1 <![CDATA[NaH2PO4]]> 1.4 Example 1-2 Example 1-2 Untreated -16.0 Example 2-3 Example 1-2 <![CDATA[KH2PO4]]> 2.9 Embodiment 2-4 Example 1-2 <![CDATA[NaH2PO4]]> -0.6

[0130] The surface area and pore width of Example 1-1, Example 2-1 and Example 2-2 were analyzed by BET. The average BET surface area and pore width are summarized in Table 6. As shown in the data in Table 6, treating the initial glass powder with an acidic phosphate solution increases the BET surface area and reduces the pore width.

[0131] Table 6

[0132]

[0133] Example 3: Nucleic acid loading on glass powder

[0134] In Example 3, nucleic acids were loaded onto the initial glass powder of Example 1 and the modified glass particles of Example 2 to form a bioactive glass carrier. Specifically, 3-phosphoglyceraldehyde dehydrogenase (GADPH) with 195 base pairs was loaded onto Examples 1-1, 2-1, 2-2, 1-2, 2-3, and 2-4 to determine DNA payload estimation and binding rate. GADPH, as an artificial DNA substitute, mimics the typical RNAi fragment length. The GADPH-loaded glass powder of Example 3 was prepared as follows: 5 mg of the glass powder of Example 1 or Example 2 (5 mg / 40 μL ethanol) was combined with a GADPH solution (6,000 ng / 60 μL H2O), mixed thoroughly by vortexing, and then incubated in a microtube equipped with a gentle shaker (300 rpm) at room temperature for 30 minutes. The sample was centrifuged at 14,000 rpm for 5 minutes to obtain a 10 μL supernatant sample for DNA quantitative analysis in the Qubit-HS-dsDNA assay. The DNA payload and binding rate for Examples 3-1 to 3-6 are recorded in Table 7. As shown in the data in Table 7, treatment of the initial glass powder with an acidic phosphate solution increased the GADPH loading on the glass powder containing the modified glass particles.

[0135] Table 7

[0136]

[0137]

[0138] Example 4: Elution of Nucleic Acids from Bioactive Glass Supports

[0139] After forming the bioactive glass carrier in embodiment 3, the elution efficiency of embodiment 3-1 to 3-4 was evaluated under condition variation. First, before quantitative analysis, the bioactive glass carrier was centrifuged to remove all supernatants. According to Table 8 and Table 9, 300 μL of phosphate buffer was added to each bioactive glass carrier of 10 mg. The solution was suspended again by vortex, and incubated at room temperature for 30 minutes using a gentle oscillator (300 rpm). The solution was centrifuged at 14,000 rpm for 5 minutes to remove all supernatants, and was used for the Qubit HS dsDNA test to carry out DNA quantitative analysis (10 μL samples were tested). The elution efficiency recorded in Table 8 and Table 9 is the percentage of the nucleic acid released (relative to the total amount of nucleic acid loaded in embodiment 3). As a control, the bioactive glass carrier was also added to water and NaCl, but no nucleic acid release was detected.

[0140] Table 8

[0141]

[0142]

[0143] Table 9

[0144] Example Glass powder Solution concentration (mM) Solution pH Elution efficiency (%) Embodiment 4-16 Example 3-3 50 5.4 0 Example 4-17 Example 3-3 50 6.4 41.3 Embodiment 4-18 Example 3-3 50 7.4 74.4 Embodiment 4-19 Example 3-3 50 8.4 51.9 Embodiment 4-20 Example 3-3 50 9.4 74.8 Example 4-21 Example 3-3 200 5.4 46.6 Example 4-22 Example 3-3 200 6.4 69.6 Example 4-23 Example 3-3 200 7.4 77.9 Example 4-24 Example 3-3 200 8.4 56.0 Example 4-25 Example 3-3 200 9.4 76.3 Example 4-26 Example 3-3 500 5.4 51.4 Example 4-27 Example 3-3 500 6.4 70.7 Embodiment 4-28 Example 3-3 500 7.4 78.7 Example 4-29 Example 3-3 500 8.4 57.8 Embodiment 4-30 Example 3-3 500 9.4 76.8

[0145] The results of nucleic acid loading in Example 4 and nucleic acid elution in Example 5 demonstrate the potential of glass powder for nucleic acid transfer and delivery. Specifically, it was observed that treating the initial glass powder with phosphate as described herein can enhance the nucleic acid loading efficiency capacity under neutral conditions. In addition, the captured nucleic acids can be released in a slightly acidic buffer system within an elution time of 30 minutes or less.

[0146] Example 5: Nuclease resistance of bioactive glass carriers

[0147] The nuclease resistance of the bioactive glass carrier of Example 3 was evaluated. Specifically, the glass powder loaded with nucleic acid of Example 3 was subjected to harsh DNase challenge conditions to simulate the attack of nucleases in certain biological states. First, before quantitative analysis, the bioactive glass carrier was centrifuged to remove all supernatants. 180 μL of H2O was added to 5 mg of the glass powder loaded with nucleic acid of Example 3, followed by 20 μL of 10X TURBO DNase buffer and 2 μL of TURBODNase (4 units, wherein 1 unit is the amount of enzyme required to decompose 1 μg of DNA within 10 minutes at 37°C). The resulting mixture was incubated at 37°C for 30 minutes. After the mixture was incubated, the mixture was heated at 75°C for 10 minutes, cooled to room temperature, and centrifuged at 15,000rpm for 10 minutes. The supernatant of each sample was removed. The sample was then rinsed with 200 μL of H2O, centrifuged at 15,500rpm for 10 minutes, and all supernatants were removed. The recovered DNA was checked by qPCR using quantitative determination. The protection rate was calculated by qPCR and is summarized in Table 10.

[0148] Table 10

[0149] Example Bioactive glass carrier Acid phosphate solution Protection rate (%) Comparative Example 5A Comparative Example 3A Untreated 3.1 Example 5-1 Example 3-1 <![CDATA[KH2PO4]]> 1.8 Example 5-2 Example 3-2 <![CDATA[NaH2PO4]]> 9.9 Comparative Example 5B Comparative Example 3B Untreated 22.6 Example 5-3 Example 3-3 <![CDATA[KH2PO4]]> 18.6 Example 5-4 Embodiment 3-4 <![CDATA[NaH2PO4]]> 27.4

[0150] The recovered DNA was also checked using integrity confirmation by electrophoresis. The DNA quantity and integrity were compared to their corresponding DNase treated and untreated samples. Figure 3 DNA input 302, DNA control 304, ladder 306, Example 3-3 308, Example 5-3 310, Example 3-4 312, Example 5-4 314, ladder 316, Example 3-1 318, Example 5-1 320, Example 3-2 322, and Example 5-2 324 are shown. Figure 3 As shown. The 195 bp related DNA fragment is Figure 3 Framed and labeled.

[0151] Example 6: Cellular Internalization of Bioactive Glass Carriers

[0152] The cell uptake efficiency and biocompatibility of the glass powders of Examples 1-2 (untreated), 2-3 (KH2PO4 treated) and 2-4 (NaH2PO4P treated) were evaluated by in vitro testing. Specifically, Examples 1-2, 2-3 and 2-4 were loaded with fluorescently labeled dsRNA to form a bioactive glass carrier. After 48 hours of co-culture with the bioactive glass carrier, fluorescently labeled dsRNA was observed in cultured insect cells sf9. As a control, the cells were cultured with dsRNA in the absence of glass powder. Figure 4 Fluorescence images and merged confocal images are shown, confirming the internalization of the bioactive glass carrier in the cell. The internalization of the dsRNA and the bioactive glass carrier can be observed under a fluorescent microscope, while no dsNRA signal was detected in the cell in the case of dsRNA alone. Since the glass powder treated with acidic phosphate solution has a higher nucleic acid payload, it is expected that a larger amount of dsRNA is carried into the target cell relative to the untreated example.

[0153] Unless otherwise expressly stated, any method described herein should not be construed as requiring that its steps be performed in a specific order, or requiring that any device have a particular orientation. Therefore, if a method claim does not actually recite the order in which its steps are to be followed, or if any device claim does not actually recite the order or orientation of the components, or if there is no other specific statement in the claims or specification that the steps are limited to a specific order, or if a specific order or orientation of the device components is not recited, then no order or orientation should be inferred in any respect. The same applies to any possible non-expressly stated basis for interpretation, including: logic regarding the setting of steps, operational flow, component order, or component orientation; ordinary meaning derived from grammatical structure or punctuation; and the number or type of aspects described in the specification.

[0154] All disclosed ranges are understood to include and provide support for claims stating any and all subranges or any and all individual values ​​contained by each range. For example, the stated range of 1 to 10 should be considered to include and provide support for any and all subranges or individual values ​​between and / or including a minimum value of 1 and a maximum value of 10; that is, all subranges start at a minimum value of 1 or greater and end at a maximum value of 10 or less (e.g., 5.5 to 10, 2.34 to 3.56, etc.), or any value from 1 to 10 (e.g., 3, 5.8, 9.9994, etc.). Any listed range can be easily identified as fully describing and allowing the same range to be decomposed into at least half, one-third, one-quarter, one-fifth, one-tenth, etc. As a non-limiting example, each range discussed herein can be easily decomposed into a lower third, a middle third, and an upper third, etc. It will also be understood by those skilled in the art that all languages, such as "up to", "at least", "greater than", "less than", etc., include the stated numbers and relate to ranges that can be subsequently decomposed into subranges discussed above. Finally, as will be understood by those skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 layers refers to groups having 1, 2, or 3 layers. Similarly, a group having 1-5 layers refers to groups having 1, 2, 3, 4, or 5 layers, and so on.

[0155] The drawings should be read as showing one or more aspects drawn to scale and / or one or more aspects not drawn to scale. This means that the drawings may be read as showing, for example, the following: (a) everything is drawn to scale; (b) nothing is drawn to scale; or (c) one or more features are drawn to scale and one or more features are not drawn to scale. Thus, the drawings may be used to provide support for the size, proportions, and / or other dimensions of any illustrated feature described individually or relative to each other. Furthermore, all such sizes, proportions, and / or other dimensions are understood to vary from 0-100% in either direction, thereby providing support for claims stating such values ​​or any and all ranges or sub-ranges that may be formed from such values.

[0156] Terms recited in the claims should be determined by reference to relevant entries in widely used general dictionaries and / or dictionaries of relevant technology, meanings commonly understood by persons skilled in the art, etc., to determine their ordinary and customary meanings, and it is understood that the terms of the claims have the broadest meaning given by any one or combination of these sources (e.g., two or more relevant dictionary entries should be combined to provide the broadest meaning of the combination of the entries, etc.), subject only to the following exceptions: (a) if a term is used in a manner broader than its ordinary and customary meaning, the term should be given its ordinary and customary meaning plus an additional extended meaning, or (b) if a term is expressly defined as having a different meaning by stating that the term is "as used in this document, the term shall mean" or similar language (e.g., "this term means," "this term is defined as," "for the purposes of this disclosure, this term shall mean," etc.). Reference to specific examples, use of "i.e.," use of the term "technology," etc. is not intended to invoke exception (b) or otherwise limit the scope of the claim terms. Except where exception (b) applies, nothing contained in this document shall be construed as a waiver of a claim or a denial of the scope of a claim.

[0157] Unless otherwise defined, all terms used herein (including technical terms and scientific terms) have the same meanings as commonly understood by those of ordinary skill in the art to which the present technology belongs. It will also be understood that terms (such as those defined in commonly used dictionaries) should be understood to be consistent with their definitions in the present application and related fields, and unless otherwise defined herein, should not be understood as idealized or fully formalized meanings. Although not clearly defined below, such terms should be interpreted according to their common meanings.

[0158] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0159] Unless the context clearly states otherwise, it is intended that the various features of the present technology described herein can be used in any combination. In addition, in some aspects, the present disclosure also contemplates that any feature or combination of features described herein can be excluded or omitted. Explanatory note, if the specification states that a consortium includes components A, B, and C, it is specifically intended that any or any combination of A, B, or C can be omitted or abandoned individually or in any combination thereof.

[0160] Unless expressly stated otherwise, all specific aspects, features and terms are intended to include both the stated aspect, feature or term and its biological equivalents.

[0161] All patents, patent applications, provisional applications, and publications referred to or cited herein are hereby incorporated by reference in their entirety, including all figures and tables, to the extent they are not inconsistent with the teachings of this specification.

Claims

1. A glass powder comprising a plurality of modified glass particles, In terms of weight percentage (wt%), the plurality of modified glass particles comprises: Greater than or equal to 20 weight percent (wt) % to less than or equal to 75 weight % SiO2; Greater than or equal to 3 mol % and less than or equal to 25 mol % P2O5; 2 wt % or more and 50 wt % or less CaO; and The total amount of brushite and apatite contained on the surfaces of the plurality of modified glass particles is greater than or equal to 30% by weight of the total weight of the modified glass particles, wherein The plurality of modified glass particles bind to the nucleic acid compound after contacting.

2. The glass powder according to claim 1, wherein The plurality of modified glass particles also include greater than 0 wt % to less than or equal to 5 wt % B 2 O 3 .

3. The glass powder according to claim 1 or 2, wherein The plurality of modified glass particles also include greater than 0 wt % to less than or equal to 5 wt % Al 2 O 3 .

4. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles also include greater than 0 wt % to less than or equal to 30 wt % Na2O.

5. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles also include greater than 0 wt % to less than or equal to 10 wt % K2O.

6. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles also include greater than 0 wt % to less than or equal to 20 wt % MgO.

7. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles also include greater than 0 wt % to less than or equal to 10 wt % ZrO 2 .

8. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles also include greater than 0 wt % to less than or equal to 5 wt % F.

9. A glass powder as claimed in any preceding claim, wherein The surfaces of the plurality of modified glass particles include brushite in an amount greater than or equal to 10 weight percent of the total weight of the modified glass particles and apatite in an amount greater than or equal to 15 weight percent of the total weight of the modified glass particles.

10. A glass powder as claimed in any preceding claim, wherein The surfaces of the plurality of modified glass particles include brushite in an amount greater than or equal to 50 weight % of the total weight of the modified glass particles and apatite in an amount greater than or equal to 3 weight % of the total weight of the modified glass particles.

11. A glass powder as claimed in any preceding claim, wherein The surfaces of the plurality of modified glass particles include a total amount of brushite and apatite greater than or equal to 60 weight % of the total weight of the modified glass particles.

12. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles include greater than or equal to 100 m 2 / g to less than or equal to 300m 2 / g of BET surface area.

13. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles comprises an average pore width of greater than or equal to 2 nanometers (nm) to less than or equal to 10 nm, as determined by BET.

14. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles comprises a zeta potential of greater than or equal to -10 mV to less than or equal to 5 mV.

15. A glass powder as claimed in any preceding claim, wherein The surfaces of the plurality of modified glass particles include needle-shaped crystals having a maximum length of greater than or equal to 10 nm and less than or equal to 50 nm.

16. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles have an average longest dimension of greater than or equal to 0.5 micrometers (μm) to less than or equal to 10 μm.

17. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles comprises: 25 wt% or more and 50 wt% or less SiO2; 8 mol % or more and 25 mol % or less P2O5; and Greater than or equal to 30 wt% to less than or equal to 50 wt% CaO.

18. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles comprises: 25 wt% or more and 35 wt% or less SiO2; 15 mol% or more and 25 mol% or less P2O5; 3 wt% or more and 6 wt% or less MgO; 38 wt% or more and 48 wt% or less CaO; as well as Greater than or equal to 2 wt% to less than or equal to 6 wt% ZrO2.

19. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles comprises: 38 wt% or more and 48 wt% or less SiO2; Greater than or equal to 5 mol % and less than or equal to 15 mol % P2O5; 10 wt% or more and 20 wt% or less MgO; and Greater than or equal to 28 wt% to less than or equal to 38 wt% CaO.

20. A glass powder as claimed in any preceding claim, wherein The plurality of modified glass particles also includes a nucleic acid compound.

21. The glass powder according to claim 20, wherein The nucleic acid compound includes: deoxyribonucleic acid (DNA) containing less than or equal to 1500 base pairs, ribonucleic acid (RNA) containing less than or equal to 1000 base pairs, or a combination thereof.

22. The glass powder according to claim 20 or 21, wherein The concentration of the nucleic acid compound is greater than or equal to 500 nanograms per milligram (ng / mg) to less than or equal to 5,000 ng / mg, based on the total weight of the plurality of modified glass particles.

23. A method of forming a glass powder, the method comprising: processing the glass into precursor glass particles, wherein the precursor glass particles have an average longest dimension of greater than or equal to 0.5 micrometers (μm) to less than or equal to 10 μm; and The precursor glass particles are treated with a treatment solution, wherein: The treatment solution comprises a phosphate; and processing to form modified glass particles; in: The surfaces of the plurality of modified glass particles include a total amount of brushite and apatite greater than or equal to 30 weight percent of the total weight of the modified glass particles; and The plurality of modified glass particles bind to the nucleic acid compound after contacting.

24. The method of claim 23, wherein: The pH of the treatment solution is greater than or equal to 2.5 and less than or equal to 4.

5.

25. The method of claim 23 or 24, wherein: The processing includes forming a mixture comprising precursor glass particles and a processing solution; and The phosphate concentration in the mixture is greater than or equal to 0.1 molar (M) to less than or equal to 1M.

26. The method of any one of claims 23 to 25, wherein: The treatment solution includes sodium sulfate, potassium sulfate, or a combination thereof.

27. The method of any one of claims 23 to 26, wherein: Processing includes: forming a mixture comprising precursor glass particles and a processing solution; and The mixture is heated at a temperature of 40°C to 80°C for the heating duration.

28. The method of claim 27, wherein: The heating duration is greater than or equal to 1 day.

29. A method of forming a bioactive glass carrier, the method comprising: processing the glass into precursor glass particles, wherein the precursor glass particles have an average longest dimension of greater than or equal to 0.5 micrometers (μm) to less than or equal to 10 μm; and The precursor glass particles are treated with a treatment solution, wherein: The treatment solution comprises a phosphate; and processing to form modified glass particles; contacting the plurality of modified glass particles with a nucleic acid compound to form a mixture; and The mixture is incubated under conditions where the nucleic acid compound non-covalently binds to the plurality of modified glass particles to form a bioactive glass carrier.

30. The method of claim 29, wherein: The mixture comprises a nucleic acid compound at a concentration of greater than or equal to 500 ng per mg of the plurality of modified glass particles (ng / mg) to less than or equal to 10,000 ng / mg.

31. The method of claim 29 or 30, wherein: The mixture is incubated at a temperature of greater than or equal to 20°C and less than or equal to 40°C.

32. The method of any one of claims 29 to 31, wherein: The mixture is incubated for a period of time greater than or equal to 10 minutes.

33. The method of any one of claims 29 to 32, wherein: Incubating the mixture includes loading the nucleic acid compound onto the plurality of modified glass particles at a loading efficiency greater than or equal to 50%.

34. The method of any one of claims 29 to 33, wherein: The bioactive glass carrier comprises a nucleic acid compound at a concentration of greater than or equal to 500 ng per mg of the bioactive glass carrier (ng / mg) to less than or equal to 5,000 ng / mg.

35. A vehicle comprising the glass powder according to any one of claims 1 to 22.

36. The carrier of claim 35, wherein The carriers include: drug delivery carriers, gene delivery carriers, tissue regeneration carriers, crop treatment or crop protection carriers, immunomodulatory carriers or gene editing complex carriers.

37. The carrier of claim 35 or 36, wherein At least a portion of the nucleic acid compound is released at a pH of 3 or more and 10 or less.

38. The carrier of claim 37, wherein At least 60 weight percent of the nucleic acid compound is released, based on the total weight of the nucleic acid compound loaded in the carrier.

39. A method for releasing a nucleic acid compound from a glass powder according to any one of claims 20-22 or 35-38, the method comprising: The glass powder is placed in a cellular environment, wherein at least a portion of the nucleic acid compound is released from the glass powder in the cellular environment.

40. The method of claim 39, wherein: At least a portion of the nucleic acid compound is released at a pH of 3 or more and 10 or less.

41. The method of claim 39 or 40, wherein: At least 60 weight percent of the nucleic acid compound is released, based on the total weight of the nucleic acid compound loaded in the glass powder.

42. The method of any one of claims 39 to 41, wherein: The cellular environment includes one or more of the following: insect cells, animal cells and embryos, fungi, fish, yeast, plant cells, or any combination thereof.

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