Bioactive glass particles as nucleic acid carriers for controlled-release

Abstract

A glass powder includes a plurality of modified glass particles, the plurality of modified glass particles comprising, in weight percent (wt.%): 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; greater than or equal to 2 wt.% to less than or equal to 50 wt.% CaO; and a surface of the plurality of modified glass particles comprises a total amount of brushite and apatite of 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 upon contact.

Description

BIOACTIVE GLASS PARTICLES AS NUCLEIC ACID CARRIERS FORCONTROLLED-RELEASECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of Chinese Patent Application Serial No. 202310986665.3, filed August 7, 2023, the entirety of which is incorporated herein by reference for all purposes.TECHNICAL FIELD

[0002] This disclosure relates to glass compositions for delivery of nucleic acids in to cells or microorganisms. In particular, the disclosure relates to bioactive glass compositions capable of capturing and releasing nucleic acids in a designated environment.BACKGROUND

[0003] Utilizing or transferring external nucleic acid for gene regulation or modulation is an emerging area of interest, especially in the agriculture and therapeutics fields. The ability of small interfering RNA (siRNA) to regulate gene expression has potential therapeutic applications, but its use is limited by inefficient delivery. The delivery of external nucleic acids into target organs or cells remains specifically challenging due to the low stability of nucleic acids and low cellular uptake efficiency. Therefore, there is a need for new glass compositions having improved surface properties which could serve as nucleic acid carriers to enhance the cell uptake rate.SUMMARY

[0004] A first aspect may be directed to a glass powder comprising a plurality of modified glass particles, the plurality of modified glass particles comprising, in weight percent (wt.%): greater than or equal to 20 weight percent (wt.%) to less than or equal to 75 wt.% SiCE, 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 surface of the plurality of modified glass particles comprises a total amount of brushite and apatite of 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 upon contact.

[0005] A second aspect of the present disclosure may include the first aspect, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 5 wt.% B2O3.

[0006] A third aspect of the present disclosure may include the first aspect or the second aspect, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 5 wt.% AI2O3.

[0007] A fourth aspect of the present disclosure may include any one of the first through third aspects, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 30 wt.% Na2O.

[0008] A fifth aspect of the present disclosure may include any one of the first through fourth aspects, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 10 wt.% K2O.

[0009] A sixth aspect of the present disclosure may include any one of the first through fifth aspects, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 20 wt.% MgO.

[0010] A seventh aspect of the present disclosure may include any one of the first through sixth aspects, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 10 wt.% ZrCh.

[0011] An eighth aspect of the present disclosure may include any one of the first through seventh aspects, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 5 wt.% F.

[0012] A ninth aspect of the present disclosure may include any one of the first through eighth aspects, wherein the surface of the plurality of modified glass particles comprises an amount of brushite of greater than or equal to 10 wt.% of the total weight of the modified glass particles and an amount of apatite of greater than or equal to 15 wt.% of the total weight of the modified glass particles.

[0013] A tenth aspect of the present disclosure may include any one of the first through ninth aspects, wherein the surface of the plurality of modified glass particles comprises an amount of brushite of greater than or equal to 50 wt.% of the total weight of the modified glass particles and an amount of apatite of greater than or equal to 3 wt.% of the total weight of the modified glass particles.

[0014] An eleventh aspect of the present disclosure may include any one of the first through tenth aspects, wherein a surface of the plurality of modified glass particles comprises a total amount of brushite and apatite of greater than or equal to 60 wt.% of the total weight of the modified glass particles.

[0015] A twelfth aspect of the present disclosure may include any one of the first through eleventh aspects, wherein the plurality of modified glass particles comprise a BET surface area of greater than or equal to 100 m2 / g to less than or equal to 300 m2 / g.

[0016] A thirteenth aspect of the present disclosure may include any one of the first through twelfth aspects, wherein the plurality of modified glass particles comprise 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.

[0017] A fourteenth aspect of the present disclosure may include any one of the first through thirteenth aspects, wherein the plurality of modified glass particles comprise a zeta-potential of greater than or equal to -10 mV to less than or equal to 5 mV.

[0018] A fifteenth aspect of the present disclosure may include any one of the first through fourteenth aspects, wherein the surface of the plurality of modified glass particles comprise acicular crystals having a largest length of greater than or equal to 10 nm to less than or equal to 50 nm.

[0019] A sixteenth aspect of the present disclosure may include any one of the first through fifteenth aspects, wherein the plurality of modified glass particles have an average longest dimension of greater than or equal to 0.5 micrometers (pm) to less than or equal to 10 pm.

[0020] A seventeenth aspect of the present disclosure may include any one of the first through sixteenth aspects, wherein the plurality of modified glass particles comprise: greater than or equal to 25 wt.% to less than or equal to 50 wt.% SiCE; 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.

[0021] An eighteenth aspect of the present disclosure may include any one of the first through seventeenth aspects, wherein the plurality of modified glass particles comprise: greater than or equal to 25 wt.% to less than or equal to 35 wt.% SiCE; 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.

[0022] A nineteenth aspect of the present disclosure may include any one of the first through eighteenth aspects, wherein the plurality of modified glass particles comprise: greater than or equal to 38 wt.% to less than or equal to 48 wt.% SiCh; 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.

[0023] A twentieth aspect of the present disclosure may include any one of the first through nineteenth aspects, wherein the plurality of modified glass particles further comprise the nucleic acid compound, thereby resulting in bioactive glass carrier.

[0024] A twenty-first aspect of the present disclosure may include the twentieth aspect, or any preceding aspect, 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 combinations thereof.

[0025] A twenty-second aspect of the present disclosure may include either one of the twentieth or the twenty-first aspects, or any preceding aspect, wherein a concentration of the nucleic acid compound is of greater than or equal to 500 nanograms per milligram (ng / mg) to less than or equal to 5,000 ng / mg, based on a total weight of the plurality of modified glass particles.

[0026] A twenty-third aspect of the present disclosure may be directed to a method of forming a glass powder, the method comprising: processing a glass into precursor glass particles, wherein the precursor glass particles have an average longest dimension of greater than or equal to 0.5 micrometers (pm) to less than or equal to 10 pm; and treating the precursor glass particles with a treatment solution, wherein: the treatment solution comprises a phosphate salt, and the treating forms modified glass particles; wherein: a surface of the plurality of modified glass particles comprises a total amount of brushite and apatite of greater than or equal to 30 wt.% of the total weight of the modified glass particles, and the plurality of modified glass particles can bind to a nucleic acid compound upon contact.

[0027] A twenty-fourth aspect of the present disclosure may include the twenty-third aspect, or any preceding aspect, wherein a pH of the treatment solution is of greater than or equal to 2.5 to less than or equal to 4.5.

[0028] A twenty-fifth aspect of the present disclosure may include either one of the twenty- third or twenty -fourth aspects, or any preceding aspect, wherein: the treating comprises forming a mixture comprising the precursor glass particles and treatment solution; and aconcentration of the phosphate salt in the mixture is greater than or equal to 0.1 molar (M) to less than or equal to 1 M.

[0029] A twenty-sixth aspect of the present disclosure may include any one of the twenty -third through twenty -fifth aspects, or any preceding aspect, wherein the treatment solution comprises sodium phosphate, potassium phosphate, or a combination thereof.

[0030] A twenty-seventh aspect of the present disclosure may include any one of the twenty- third through twenty-sixth aspects, or any preceding aspect, wherein the treating comprises: forming a mixture comprising the precursor glass particles and the treatment solution; and heating the mixture at a temperature of from 40 °C to 80 °C for a heating duration.

[0031] A twenty-eighth aspect of the present disclosure may include the twenty- seventh aspect, or any preceding aspect, wherein the heating duration is greater than or equal to 1 day.

[0032] A twenty-ninth aspect of the present disclosure may be directed to a method of forming a bioactive glass carrier, the method comprising: processing a glass into precursor glass particles, wherein the precursor glass particles have an average longest dimension of greater than or equal to 0.5 micrometers (pm) to less than or equal to 10 pm; and treating the precursor glass particles with a treatment solution, wherein: the treatment solution comprises a phosphate salt, and the treating forms modified glass particles; contacting the plurality of modified glass particles with a nucleic acid compound to form an admixture; and incubating the admixture under conditions in which the nucleic acid compound non-covalently binds to the plurality of modified glass particles, forming the bioactive glass carrier.

[0033] A thirtieth aspect of the present disclosure may include the twenty-ninth aspect, or any preceding aspect, wherein the admixture comprises a concentration of the nucleic acid compound 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.

[0034] A thirty -first aspect of the present disclosure may include either one of the twenty -ninth or the thirtieth aspects, or any preceding aspect, wherein the incubating the admixture is conducted at a temperature of greater than or equal to 20 °C to less than or equal to 40 °C.

[0035] A thirty-second aspect of the present disclosure may include any one of the twentyninth through thirty-first aspects, or any preceding aspect, wherein the incubating the admixture is conducted for a period greater than or equal to 10 minutes.

[0036] A thirty -third aspect of the present disclosure may include any one of the twenty -ninth through thirty-second aspects, or any preceding aspect, wherein the incubating the admixture comprises loading the nucleic acid compound on the plurality of modified glass particles at a loading efficiency of greater than or equal to 50%.

[0037] A thirty-fourth aspect of the present disclosure may include any one of the twenty -ninth through thirty-third aspects, or any preceding aspect, wherein the bioactive glass carrier comprises a concentration of the nucleic acid compound 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.

[0038] A thirty-fifth aspect of the present disclosure may be directed to a vehicle comprising the glass powder (or a bioactive glass carrier) according to any one of the first through twenty- second aspects, or any preceding aspect.

[0039] A thirty-sixth aspect of the present disclosure may include the thirty-fifth aspect, or any preceding aspect, wherein the vehicle comprises a drug delivery vehicle, a gene delivery vehicle, a tissue regeneration vehicle, a crop treatment or crop protection vehicle, an immunomodulatory vehicle, or a gene editing complex vehicle.

[0040] A thirty-seventh aspect of the present disclosure may include either one of the thirtyfifth or the thirty-sixth aspects, or any preceding aspect, wherein at least a portion of the nucleic acid compound is released at a pH greater than or equal to 3 to less than or equal to 10.

[0041] A thirty-eighth aspect of the present disclosure may include the thirty-seventh aspect, or any preceding aspect, wherein at least 60 weight percent of the nucleic acid compound is released, based on a total weight of the nucleic acid compound loaded in the vehicle.

[0042] A thirty-ninth aspect of the present disclosure may be directed to a method of releasing the nucleic acid compound from the glass powder (or bioactive glass carrier) of any one of the twentieth through twenty-second or thirty-fifth through thirty-eighth aspects, or any preceding aspect, the method comprising: positioning the glass powder (or bioactive glass carrier) in a cellular environment, wherein at least a portion of the nucleic acid compounds is released from the glass powder (or bioactive glass carrier) in the cellular environment.

[0043] A fortieth aspect of the present disclosure may include the thirty-ninth aspect, or any preceding aspect, wherein at least a portion of the nucleic acid compound is released at a pH greater than or equal to 3 to less than or equal to 10.

[0044] A forty-first aspect of the present disclosure may include the thirty-ninth or fortieth aspect, or any preceding aspect, wherein at least 60 weight percent of the nucleic acid compound is released, based on a total weight of the nucleic acid compound in the glass powder (or bioactive glass carrier).

[0045] A forty-second aspect of the present disclosure may include any one of the thirty-ninth through forty-first aspects, or any preceding aspect, 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

[0046] FIG. 1 provides X-ray diffraction (XRD) patterns with a plot of intensity (y-axis) as a function of position (x-axis) for the modified glass particles in accordance with one or more aspects shown and described herein;

[0047] FIG. 2A provides a scanning electron microscopy (SEM) image of precursor glass particles in accordance with one or more aspects shown and described herein;

[0048] FIG. 2B provides a scanning electron microscopy (SEM) image of modified glass particles in accordance with one or more aspects shown and described herein;

[0049] FIG. 2C provides a scanning electron microscopy (SEM) image of modified glass particles in accordance with one or more aspects shown and described herein;

[0050] FIG. 2D provides a scanning electron microscopy (SEM) image of precursor glass particles in accordance with one or more aspects shown and described herein;

[0051] FIG. 2E provides a scanning electron microscopy (SEM) image of modified glass particles in accordance with one or more aspects shown and described herein;

[0052] FIG. 2F provides a scanning electron microscopy (SEM) image of modified glass particles in accordance with one or more aspects shown and described herein;

[0053] FIG. 3 provides electrophoresis images of glass powders subjected to DNase challenging in accordance with one or more aspects shown and described herein; and

[0054] FIG. 4 provides fluorescence and merged confocal images of glass powders internalized in cells in accordance with one or more aspects shown and described herein.

[0055] FIG. 5 provides a graph showing larvae survival rate over time for modified glass particles in accordance with one or more aspects shown herein as compared to positive and negative controls.DETAILED DESCRIPTION

[0056] Reference will now be made in detail to glass particles which may be used as nucleic acid carriers, various aspects which are illustrated in the accompanying drawings. Whenever possible, the same reference numerals will be used throughout the drawings to refer to the same or like parts. According to aspects, a glass powder may comprise a plurality of modified glass particles which may be used as nucleic acid carriers. The plurality of modified glass particles may include, in weight percent (wt.%): greater than or equal to 20 weight percent (wt.%) to less than or equal to 75 wt.% SiCh; 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. A surface of each of the plurality of modified glass particles may include a total amount of brushite and apatite of greater than or equal to 30 wt.% of the total weight of the modified glass particles, wherein the modified glass particles bind to a nucleic acid compound upon contact. Various aspects of glass particles for use as nucleic acid carriers will be described herein with specific reference to the drawings. The components in the drawings are not necessarily to scale, emphasis instead being placed upon illustrating the principles of the exemplary aspects.

[0057] The present disclosure generally relates to glass powders, methods of forming glass powders, and methods of forming bioactive glass carriers.

[0058] The definitions of certain terms used in the present disclosure are provided below. Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which the present technology belongs.

[0059] As used in this disclosure and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to “a” component includes aspects having two or more such components, unless the context clearly indicates otherwise. For example, reference to “a cell” includes a combinationof two or more cells, and the like. Also, the word “or” when used without a preceding “either” (or other similar language indicating that “or” is unequivocally meant to be exclusive - e.g., only one of x or y, etc.) shall be interpreted to be inclusive (e.g., “x or y” means one or both x or y). Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, analytical chemistry and nucleic acid chemistry described below are those well-known and commonly employed in the art.

[0060] The term “and / or” shall also be interpreted to be inclusive (e.g., “x and / or y” means one or both x or y). In situations where “and / or” or “or” are used as a conjunction for a group of three or more items, the group should be interpreted to include one item alone, all the items together, or any combination or number of the items. Moreover, terms used in the specification and claims such as have, having, include, and including should be construed to be synonymous with the terms comprise and comprising. Other elements may optionally be present other than the elements specifically identified by the “and / or” clause, whether related or unrelated to those elements specifically identified. As a non-limiting example, a reference to “X and / or Y” can refer, in one aspect, to X only (optionally including elements other than Y); in some aspects, to Y only (optionally including elements other than X); in yet some aspects, to both X and Y (optionally including other elements).

[0061] As used herein, the term "particle size" refers to the longest dimension of the particle, unless otherwise stated. The “average particle size” refers to an average of the longest dimension of a plurality of the particles. In aspects where the particle is spherical, the longest dimension is the particle diameter. The particle size is expressed, for example, as an average particle size or average longest dimension of a plurality of the particles, which may be determined using electron microscopy in conjunction with image analysis.

[0062] As used herein the terms "bioactive glass" (“BAG”) or "biologically active glass" means an inorganic glass material which is capable of binding with desired biological material, such as nucleic acids. As used herein, the term “bioactive glass carrier” means a glass material which is bound with desired biological material, such as a nucleic acid.

[0063] As used herein, the term "controlled release" refers to release of the nucleic acid from a bioactive glass carrier according to a predetermined profile. In some aspects, the selected controlled release may include the release of the nucleic acid compound in a particular pH range, ionic strength range, or both.

[0064] Various aspects and / or embodiments of this disclosure relate to glass powders including modified glass particles. In various aspects, a surface of the modified glass particle includes a total amount of brushite and apatite greater than or equal to 30 wt.% of the total weight of the modified glass particle. The modified glass particles are capable of binding to nucleic acids upon contact. The glass powders disclosed herein exhibit excellent biocompatibility. The bioactive nature of the glass powder results from a series of reactions on the surface of precursor glass particles under certain conditions. For example, when the precursor glass particles are exposed to a treatment solution including phosphate, cation exchange may occur, such that the cations from the glass surface are replaced by cations from the treatment solution. Ca2+ions released from the glass surface due to the dissolution of the glass network in combination with PO43ions in solution can react on the glass surface. This favors the formation of brushite (CaHPO4 2H2O), a known bioactive ceramic, or apatite, such as hydroxyapatite (Caio(P04)e(OH)2). Both brushite and apatite are capable of binding to nucleic acids, such as DNA and RNA. In aspects, the apatite may refer to hydroxyapatite, fluoroapatite (Cas(PO4)3F), chloroapatite (Ca5(PO4)3F), or combinations thereof. In aspects, the apatite may refer to hydroxyapatite.

[0065] A glass powder may be formed comprising a plurality of modified glass particles. In aspects, the modified glass particles of the glass powder are formed from precursor glass particles. The glass powder may exhibit excellent nucleic acid capturing / releasing efficiency.

[0066] A method of forming the glass powder may comprise processing a glass into precursor glass particles. Depending on the composition, glasses can be melted at a temperature in the range of about 1000 °C to about 1600 °C. For example, glasses can be melted at a temperature in the range of about 1100 °C to about 15000C, or even at a temperature in the range of about 1150 °C to about 1450° C. The glasses may be melted in a suitable apparatus, e.g., a covered platinum crucible, for a time sufficient to melt the glasses, such as e.g., about 60 minutes (min) to about 24 hours (h), including from about 90 min to about 20 h, about 2 h to about 18 h, about 5 h to about 16 h, about 8 h to about 12 h, or any range including and / or in between any two of these values. After melting and fining, the glasses can be poured and annealed at a temperature of about 350 °C to about 550 °C, such as about 400 °C to about 500 °C. The annealed glasses can then be cast in to a suitable mold, following which the resulting materials can be cooled to form the glass. In certain aspects, the glass may be ground into precursor glass particles having particle sizes in the range of from 0.5 to 10 pm by air jet milling. The particle size can be variedin the range of 1-100 gm using attrition milling or ball milling. Furthermore, these glasses can be subsequently processed into submicron particle sizes using different methods, such as planetary ball milling or wet milling. In aspects, the precursor glass particles have an average longest dimension (particle size) of greater than or equal to 0.5 micrometers (pm) to less than or equal to 10.0 pm, such as from 0.5 pm to 9.0 pm, from 0.6 pm to 8.0 pm, from 0.7 pm to 7.0 pm, from 0.8 pm to 6.0 pm, from 1.0 pm to 5.0 pm, from 2.0 pm to 4.0 pm, or any range including and / or in between any two of these values. On an industrial scale, the precursor glass particles of desired size may be produced using the top-down melting approach followed by milling or grinding, which affords flexibility of composition design and low cost of manufacturing. In certain aspects, the glass may be ground into precursor glass particles using air jet, ball milling or grinding. The precursor glass particles of desired size can be further treated to form modified glass particles.

[0067] The method of forming the glass powder may comprise treating the precursor glass particles with a treatment solution, wherein the treatment solution comprises a phosphate salt, to form modified glass particles. In aspects, the treatment solution comprises a phosphate salt, such as but not limited to sodium phosphate, potassium phosphate, or combinations thereof. In aspects, the treatment solution is an aqueous solution.

[0068] In aspects, a pH of the treatment solution may be from 2.5 to 5.0, such as from 2.5 to 4.8, from 3.0 to 4.5, from 3.5 to 4.5, or any range including and / or in 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 of less than 2.5 may dissolve at least portions of the precursor glass particles. Further it is believed that treating the precursor glass particles with a treatment solution having a pH of greater than 5.0 may reduce apatite formation.

[0069] The treating may comprise forming a mixture comprising the precursor glass particles and the treatment solution. In aspects, a concentration of the phosphate salt in the mixture may be from 0. 1 molar (M) to 1 M, such as from 0.2 M to 0.8 M, from 0.3 M to 0.7 M, from 0.4 M to 0.6 M, or any range including and / or in between any two of these values.

[0070] In aspects, the treating may comprise heating the mixture at an elevated temperature, such as from 40 °C to 80 °C or from 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 greaterthan or equal to 5 days. In aspects, the heating duration is from 1 day to 60 days, from 1 day to 30 days, from 1 day to 15 days, from 1 day to 7 days, from 3 days to 60 days, from 3 days to 30 days, from 3 days to 15 days, from 3 days to 10 days, or from 3 days to 7 days. Without intending to be bound by any particular theory, it is believed that a heating duration of less than 1 day may not provide sufficient formation of apatite and brushite on the surface of the modified glass particles.

[0071] The treating of the precursor glass particles with the treatment solution may form modified glass particles, the modified glass particles having an increased concentration of brushite and apatite at a surface of the modified glass particles, relative to a concentration of brushite and apatite at a surface of the precursor glass particles. Further, the treating may increase a surface area of the modified glass particles. An increase in both the surface area and the brushite and apatite concentration of the modified glass particles may increase a nucleic acid loading capacity of the modified glass particles.

[0072] In aspects, a method of forming a bioactive glass carrier may comprise contacting the modified glass particles with a nucleic acid compound to form an admixture, and incubating the admixture under conditions in which the nucleic acid compound non-covalently binds to the modified glass particles, forming the bioactive glass carrier. In aspects, the admixture may include a suitable liquid medium such as water, alcohols or hydrocarbons.

[0073] The admixture may comprise a concentration of the nucleic acid compound of from 500 nanograms (ng) per milligram (mg) of the modified glass particles (ng / mg) to 10,000 ng / mg. For example, the admixture may comprise a concentration of the nucleic acid compound of from 500 ng / mg to 10,000 ng / mg, from 750 ng / mg to 8,000 ng / mg, from 1,000 ng / mg to 6,000 ng / mg, from 1,250 ng / mg to 4,000 ng / mg, from 1,500 ng / mg to 3,000 ng / mg, or any range including and / or in between any two of these values.

[0074] In aspects, incubating the admixture may comprise treating the admixture at a temperature of from 20 °C to 40 °C, such as from 20 °C to 30 °C, or from 20 °C to 25 °C. Without intending to be bound by any particular theory, it is believed that incubating the admixture a temperature of greater than 40 °C may destabilize the nucleic acid. The incubating may be conducted for a period greater than or equal to 10 minutes. In aspects, the incubating may be conducted for a period less than or equal to 3 hours, less than or equal to 2 hours, or less than or equal to 1 hour.

[0075] The loading efficiency of loading the nucleic acid on the modified glass particles can be determined by calculating an amount of the nucleic acid in the admixture (total nucleic acid added) and the residual amount of the nucleic acid in a supernatant of the admixture after removing the modified glass particles (nucleic acid not loaded). The amount of nucleic acid loaded can be determined by subtracting the residual amount of the nucleic acid in the supernatant from the total amount of nucleic acid in the admixture. The loading efficiency is calculated by dividing the amount of nucleic acid loaded by the total amount of nucleic acid in the admixture. Conventional methods in the art, such as using a microvolume ultraviolet-visible spectrophotometer or flurometric Quibit nucleic acid assay, may be used to determine a concentration of the nucleic acid in the admixture and supernatant. The surface charge of the modified glass particles and capacity may affect the loading efficiency. In aspects, incubating the admixture comprises loading the nucleic acid compound on the modified glass particles at a loading efficiency of greater than or equal to 50%, such as greater than or equal to 70%, greater than or equal to 80%, or even greater than or equal to 90%. In aspects, the loading efficiency may be from 50% to 100%, such as from 60% to 95%, from 70% to 95%, from 80% to 95%, or from 90% to 95%.

[0076] As discussed herein, the glass powder may comprise 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 influenced by the composition of the modified glass particles.

[0077] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom include SiCh. When the content of SiCh is too small, the glass may have poor chemical and mechanical durability. On the other hand, when the content of SiO? is too large, the melting ability of the glass may decrease since the melting temperature of pure SiO? is relatively high. With increasing SiO? content, the bioactivity of modified glass particles may decrease. In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom include a silicate glass, which includes SiO? as the primary glass-forming oxide. In aspects, SiO? is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount of from 20 wt. % to 75 wt. %, such as from 25 wt. % to 70 wt. %, from 30 wt. % to 65 wt. %, from 35 wt. % to 60 wt. %, from 40 wt. % to 55 wt. %, from 45 wt. % to 50 wt. % of the total weight of the precursor glass particles and modified glass particles, respectively, or any range including and / or in betweenany two of these values based on a total weight of the precursor glass particles or modified glass particles.

[0078] The precursor glass particles or the modified glass particles of the glass powder formed therefrom may also include P2O5, which serves as a network former. Furthermore, the liberation of phosphate ions to the surface of the precursor glass particles may contribute to the formation of calcium-phosphorous (Ca-P) compounds such as brushite or apatite. In addition, P2O5 may increase the viscosity of the glass, which in turn expands the range of operating temperatures, and may be an advantage to the manufacture and the formation of the glass which is subsequently processed into glass powder. In various aspects, P2O5 is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount from 3 wt. % to 25 wt. % of the total weight of the precursor glass particles or modified glass particles, such as from 8 wt. % to 25 wt.%, 15 wt.% to 25 wt. %, or from 5 wt.% to 15 wt.% of P2O5. In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom include from 3 wt. % to 25 wt. %, from 5 wt.% to 20 wt.%, from 8 wt.% to 15 wt.%, from 10 wt.% to 12 wt.%, or any range including and / or in between any two of these values of P2O5, based on the total weight of the precursor glass particles or modified glass particles.

[0079] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may include divalent cation oxides (such as 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. Further, CaO may react with a treatment solution including a phosphate salt to form apatite. The release of Ca2+ions from the surface of the precursor glass particles may contribute to the formation of a layer of the surface of the particle rich in calcium phosphate. In aspects, CaO is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount from 2 wt. % to 50 wt. %, based on the total weight of the precursor glass particles or modified glass particles. For example, in aspects, CaO is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount including from 30 wt. % to 50 wt. %, from 28 wt.% to 38 wt.%, or from 38 wt.% to 48 wt.% CaO, based on the total weight of the precursor glass particles or modified glass particles. In aspects, CaO is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount from 2 wt. % to50 wt. %, from 5 wt. % to 45 wt. %, from 10 wt. % to 40 wt. %, from 15 wt. % to 35 wt. %, from 20 wt. % to 30 wt. %, from 28 wt.% to 38 wt.%, from 38 wt.% to 48 wt.%, or any range including and / or in between any two of these values, based on the total weight of the precursor glass particles or modified glass particles.

[0080] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may include B2O3, which may increase the bioactivity of the modified glass particles of the glass powder. In aspects, B2O3 is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount from 0 wt. % to 5 wt. %, based on the total weight of the precursor glass particles or modified glass particles. For example, in aspects, B2O3 is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount including from 0 wt. % to 5 wt. %, from 0.2 wt. % to 4.5 wt. %, from 0.5 wt. % to 4 wt. %, from 1 wt. % to 3.5 wt. %, from 1.5 wt. % to 3 wt. %, from 2 wt. % to 2.5 wt. %, or any range including and / or in between any two of these values, based on the total weight of the precursor glass particles or modified glass particles.

[0081] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may include AI2O3, which may increase the thermal stability of the glass. In aspects, AI2O3 is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount from 0 wt. % to 5 wt. %, based on the total weight of the precursor glass particles or modified glass particles. For example, in aspects, AI2O3 is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount including from 0 wt. % to 5 wt. %, from 0.2 wt. % to 4.5 wt. %, from 0.5 wt. % to 4 wt. %, from 1 wt. % to 3.5 wt. %, from 1.5 wt. % to 3 wt. %, from 2 wt. % to 2.5 wt. %, or any range including and / or in between any two of these values, based on the total weight of the precursor glass particles or modified glass particles.

[0082] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may include Na2O, which may increase the bioactivity of the glass. In aspects, Na2O is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount from 0 wt. % to 30 wt. %, based on the total weight of the precursor glass particles or modified glass particles. For example, in aspects, Na2O is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount including from 0 wt. % to 30 wt. %, from 5 wt. % to25 wt. %, from 10 wt. % to 20 wt. %, from 12 wt. % to 15 wt. %, or any range including and / or in between any two of these values, based on the total weight of the precursor glass particles or modified glass particles.

[0083] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may include K2O, which may increase the bioactivity of the glass. In aspects, K2O is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount from 0 wt. % to 10 wt. %, based on the total weight of the precursor glass particles or modified glass particles. For example, in aspects, K2O is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount including from 0 wt. % to 10 wt. %, from 1 wt. % to 9 wt. %, from 2 wt. % to 8 wt. %, from 3 wt. % to 7 wt. %, from 4 wt. % to 6 wt. %, from 4 wt. % to 5 wt. %, or any range including and / or in between any two of these values, based on the total weight of the precursor glass particles or modified glass particles.

[0084] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may include MgO, which may increase the bioactivity of the glass. In aspects, MgO is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount from 0 wt. % to 20 wt. %, based on the total weight of the precursor glass particles or modified glass particles. For example, in aspects, MgO is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount including from 0 wt. % to 20 wt. %, from 1 wt. % to 18 wt. %, from 2 wt. % to 16 wt. %, from 3 wt. % to 15 wt. %, from 5 wt. % to 15 wt. %, from 6 wt.% to 14 wt.%, from 7 wt.% to 13 wt.%, from 8 wt.% to 12 wt.%, from 9 wt.% to 11 wt.%, from 10 wt.% to 20 wt.%, from 3 wt. % to 6 wt. %, or any range including and / or in between any two of these values, based on the total weight of the precursor glass particles or modified glass particles.

[0085] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may include ZrCh, which may increase the thermal stability of the glass. In aspects, ZrCh is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount from 0 wt. % to 10 wt. %, based on the total weight of the precursor glass particles or modified glass particles. For example, in aspects, ZrCh is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount including from 0 wt. % to 10 wt. %, from 1 wt. % to 9wt. %, from 2 wt. % to 8 wt. %, from 3 wt. % to 7 wt. %, from 4 wt. % to 6 wt. %, from 4 wt.% to 5 wt. %, from 2 wt. % to 6 wt. %, or any range including and / or in between any two of these values, based on the total weight of the precursor glass particles or modified glass particles.

[0086] In aspects, the precursor glass particles or the modified glass particles of the glass powder formed therefrom may include fluoride (F‘), which may increase the bioactivity of the glass. In aspects, F’ is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount from 0 wt. % to 5 wt. %, based on the total weight of the precursor glass particles or modified glass particles. For example, in aspects, F’ is present in the precursor glass particles or the modified glass particles of the glass powder formed therefrom in an amount including from 0 wt. % to 5 wt. %, from 0.2 wt. % to 4.5 wt. %, from 0.5 wt. % to 4 wt. %, from 1 wt. % to 3.5 wt. %, from 1.5 wt. % to 3 wt. %, from about 2 wt. % to about 2.5 wt. %, or any range including and / or in between any two of these values, based on the total weight of the precursor glass particles or modified glass particles.

[0087] In aspects, the modified glass particles comprise greater than or equal to 25 wt.% to less than or equal to 50 wt.% SiCh; 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.

[0088] In aspects, the modified glass particles comprise greater than or equal to 25 wt.% to less than or equal to 35 wt.% SiCh; 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.% ZrCh.

[0089] In aspects, the modified glass particles comprise greater than or equal to 38 wt.% to less than or equal to 48 wt.% SiCh; 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.

[0090] The modified glass particles formed from the precursor glass particles may exhibit certain properties that differ from the precursor glass particles, such as but not limited to brushite and apatite surface concentration, BET surface area, average pore width, or surface charge.

[0091] A concentration of brushite and / or apatite on the surface of the modified glass particles may be determined by quantitative phase analysis of XRD using the Rietveld method Rietveld, as known in the art.

[0092] In aspects, the surface of the modified glass particles may comprise an amount of brushite of greater than or equal to 5 wt.% of the total weight of the modified glass particles. For example, the surface of the modified glass particles may comprise an amount of brushite of greater than or equal to 10 wt.%, greater than or equal to 20 wt.%, greater than or equal to 30 wt.%, greater than or equal to 40 wt.%, greater than or equal to 50 wt.%, greater than or equal to 60 wt.%, or even greater than or equal to 70 wt.% of the total weight of the modified glass particles. The modified glass particles may comprise an amount of brushite of from 5 wt.% to 90 wt.%, from 10 wt.% to 80 wt.%, from 20 wt.% to 70 wt.%, from 30 wt.% to 50 wt.% of the total weight of the modified glass particles, or any range including and / or in between any two of these values.

[0093] In aspects, the surface of the modified glass particles may comprise an amount of apatite of greater than or equal to 2 wt.% of the total weight of the modified glass particles. For example, the surface of the modified glass particles may comprise an amount of apatite of greater than or equal to 3 wt.%, greater than or equal to 4 wt.%, greater than or equal to 5 wt.%, greater than or equal to 8 wt.%, or greater than or equal to 10 wt.% of the total weight of the modified glass particles. The modified glass particles may comprise an amount of apatite of from 2 wt.% to 40 wt.%, from 3 wt.% to 30 wt.%, from 4 wt.% to 20 wt.%, from 5 wt.% to 15 wt.% of the total weight of the modified glass particles, or any range including and / or in between any two of these values.

[0094] In aspects, the surface of the modified glass particles may comprise a total amount of brushite and apatite of greater than or equal to 30 wt.% of the total weight of the modified glass particles. For example, the surface of the modified glass particles may comprise a total amount of brushite and apatite of greater than or equal to 35 wt.%, greater than or equal to 40 wt.%, greater than or equal to 50 wt.%, greater than or equal to 60 wt.%, greater than or equal to 70 wt.%, or even greater than or equal to 80 wt.% of the total weight of the modified glass particles. The modified glass particles may comprise a total amount of brushite and apatite of from 30 wt.% to 95 wt.%, from 35 wt.% to 90 wt.%, from 40 wt.% to 85 wt.%, from 45 wt.% to 80 wt.%, from 50 wt.% to 75 wt.%, from 55 wt.% to 70 wt.%, from 60 wt.% to 65 wt.% ofthe total weight of the modified glass particles, or any range including and / or in between any two of these values.

[0095] In certain aspects, the surface of the modified glass particles may comprise an amount of brushite of greater than or equal to 10 wt.% of the total weight of the modified glass particles and an amount of apatite of greater than or equal to 15 wt.% of the total weight of the modified glass particles. In other aspects, the surface of the modified glass particles may comprise an amount of brushite of greater than or equal to 50 wt.% of the total weight of the modified glass particles and an amount of apatite of greater than or equal to 3 wt.% of the total weight of the modified glass particles.

[0096] In aspects, the weight percentages of apatite disclosed herein may also refer to a weight percentage of hydroxyapatite. It should be understood that all ranges and subranges regarding an amount of apatite on the surface of the modified glass particles are also disclosed for an amount of hydroxyapatite on the surface of the modified glass particles.

[0097] Without intending to be bound by any particular theory, it is believed that the modified glass particles having an increased amount of brushite and / or apatite on the surface of the modified glass particles may increase the nucleic acid loading capacity of the modified glass particles.

[0098] The modified glass particles may comprise a BET surface area of from 100 m2 / g to 300 m2 / g, such as from 100 m2 / g to 250 m2 / g, from 100 m2 / g to 200 m2 / g, from 125 m2 / g to 300 m2 / g, from 125 m2 / g to 250 m2 / g, from 125 m2 / g to 200 m2 / g, from 150 m2 / g to 300 m2 / g, from 150 m2 / g to 250 m2 / g, from 150 m2 / g to 200 m2 / g, or any range including and / or in between any two of these values. As used herein, “BET surface area” refers to the surface area, as determined by Brunauer-Emmett-Teller theory. The surface area of the modified glass particles can also be varied by changing the particle size. Without intending to be bound by any particular theory, it is believed that the modified glass particles having an increased BET surface area may increase the nucleic acid loading capacity of the modified glass particles.

[0099] The modified glass particles may comprise an average pore width of from 2 nm to 10 nm, such as from 3 nm to 9 nm, from 4 nm to 8 nm, from 5 nm to 7 nm, or any range including and / or in between any two of these values, as determined by BET.

[0100] The modified glass particles may comprise a zeta-potential of greater than or equal to - 10 mV to less than or equal to 5 mV, such as from -8 mV to 4 mV, from -5 mV to 4 mV, from -2 mV to 3 mV, or any range including and / or in between any two of these values. Without intending to be bound by any particular theory, it is believed that a surface charge of near neutral (where neutral is 0 mV zeta-potential), such as the zeta-potential ranges described herein, demonstrates less active oxide exposure of the modified glass particles. The zetapotential may be measured using conventional means in the art, such as electrophoretic light scattering (Malvern Zetasizer Nano ZS).

[0101] In aspects, the modified glass particles may comprise acicular crystals of apatite and / or brushite having a largest length of greater than or equal to 10 nm to less than or equal to 50 nm. Without intending to be bound by any particular theory, it is believed that acicular crystals on the surface of the modified glass particles may provide nano-sized pores for nucleic acid binding, which may increase the nucleic acid loading capacity of the modified glass particles. The presence of acicular crystals and the largest length of the acicular crystals may be determined by conventional means in the art, such as electron microscopy.

[0102] In aspects, the modified glass particles may have an average longest dimension of greater than or equal to 0.5 micrometers (pm) to less than or equal to 10.0 pm, such as from 0.5 pm to 9.0 pm, from 0.6 pm to 8.0 pm, from 0.7 pm to 7.0 pm, from 0.8 pm to 6.0 pm, from 1.0 pm to 5.0 pm, from 2.0 pm to 4.0 pm, or any range including and / or in between any two of these values. The average longest dimension of the modified glass particles may be determined by conventional means in the art, such as electron microscopy in conjunction with image analysis.

[0103] As described herein, the modified glass particles may bind to a nucleic acid compound upon contact. In aspects, the modified glass particles may comprise the nucleic acid compound. The nucleic acid compound may comprise DNA comprising less than or equal to 1500 base pairs, RNA comprising less than or equal to 1000 base pairs, or combinations thereof. The modified class particles comprising the nucleic acid compound may be referred to as a bioactive glass carrier.

[0104] In aspects, the bioactive glass carrier may comprise a concentration of the nucleic acid compound of from 500 ng per mg of the bioactive glass carrier (ng / mg) to 5,000 ng / mg. For example, the bioactive glass carrier may comprise a concentration of the nucleic acidcompound of from 500 ng / mg to 5,000 ng / mg, from 750 ng / mg to 3,000 ng / mg, from 1,000 ng / mg to 2,500 ng / mg, from 1,500 ng / mg to 2,000 ng / mg, or any range including and / or in between any two of these values.

[0105] The modified glass particles may bind to a nucleic acid compound under various pH conditions. The nucleic acid compound may coat or be located on the surface of the bioactive glass particle. Suitable nucleic acids include any ribonucleic acid (RNA) or deoxyribonucleic acid (DNA), which may be unmodified or modified. In various aspects, nucleic acid compound includes, without limitation, single- and double-stranded RNA, single- and double- stranded DNA, RNA that is mixture of single- and double-stranded regions, DNA that is a mixture of single- and double-stranded regions, and hybrid molecules comprising DNA and RNA that may be single-stranded or, double-stranded or a mixture of single- and double-stranded regions. In addition, the nucleic acid compound may refer to triple-stranded regions comprising RNA or DNA or both, DNAs or RNAs containing one or more modified bases, and DNAs or RNAs with backbones modified for stability or for other reasons. In some aspects, the nucleic acid compound includes ribonucleic acid, deoxyribonucleic acid, or derivatives thereof. In some aspects, the nucleic acid compound includes ribonucleic acid or derivatives thereof. In other aspects, the nucleic acid compound includes deoxyribonucleic acid or derivatives thereof. In some aspects, the nucleic acid compound includes interfering ribonucleic acid molecules.

[0106] Some properties to substantiate the capability and capacity of nucleic acid loading on the modified glass particles may be, in addition to the apatite and brushite concentration, surface charge and surface area. The surface charge can be directly determined by zeta potential measurement. The more the positive charge on the particle surface, the higher the potency to capture the negative charge of the nucleic acid compound. On the other hand, the nucleic acid compound capture and release may be controllable and reversible. The carried nucleic acid may be released while delivered into a target. It is known that the pH value inside cells is slightly acidic compared to the body fluids. Hence, the negative surface charge of carriers in acidic solution may assist nucleic acid releasing. The binding of nucleic acid on the surface of the modified glass particles may be assessed using the change in zeta-potential of the modified glass particle surface. A change in zeta-potential may be indicative of the ligand binding to the surface of the modified glass particle. The zeta-potential is a measure of surface charge on the modified glass particles and may have an impact on the stability of particles under various pH conditions.

[0107] Another aspect of the present disclosure relates to a method of releasing the nucleic acid compounds from the glass powder (e.g., bioactive glass carrier) or vehicle comprising the glass powder (or bioactive glass carrier) described herein. In aspects, the method may include positioning the glass powder (or bioactive glass carrier) in a cellular environment, wherein at least portion of the nucleic acid compounds is released from the glass powder (or bioactive glass carrier) in the cellular environment. In aspects, the cellular environment may have a pH of from 3 to 10, such as from 4 to 9, from 5 to 8, from 6 to 7, or any range including and / or in between any two of these values, or any other pH value or condition disclosed herein.

[0108] In aspects, greater than or equal to 60 wt.% of the nucleic acid compound may be released, such as greater than or equal to 65 wt.%, greater than or equal to 70 wt.%, greater than or equal to 75 wt.%, or greater than or equal to 80 wt.%, based on the total weight of the nucleic acid compound loaded in the glass powder (or bioactive glass carrier). The released nucleic acid compounds in the cellular environment may be delivered to target cells in the cellular environment. Without intending to be bound by any particular theory, it is believed that the controlled release of the nucleic acid compounds from the glass poweder (or bioactive glass carrier) in the cellular environment may selectively increase a concentration of the nucleic acid compounds in or near targeted cells in the cellular environment, thereby increasing efficiency of delivery of the nucleic acid compounds to a desired cellular environment. In some aspects, the cellular environment comprises one or more of insect cells, animal cells and embryos, fungi, fish, yeast, plant cells, or any combination thereof.

[0109] Another aspect of the present disclosure relates to a vehicle comprising the glass powder comprising the modified glass particles described herein. The vehicle may include a drug delivery vehicle, a gene delivery vehicle, a tissue regeneration vehicle, a crop treatment or crop protection vehicle, an immunomodulatory vehicle, or a gene editing complex vehicle. The vehicle can be used to deliver the modified glass particles and / or bioactive glass carriers described herein to various types of cells. Any type of cell which is susceptible to delivery of nucleic acids may be used as the target for transformation according to the present disclosure. Examples include, without limitation insect cells, animal cells and embryos, fungi, fish, yeast, and plant cells. A stable aqueous suspension may be used as part of phytosanitary compositions for controlling pest. For example, the stable suspensions may be used to protect crops against insects, in which case the nucleic acid may be insecticidal double stranded RNA. The vehicles may be used in the prevention and / or control of pest infestation of plants, using interferingribonucleic acid (RNA) molecules. Compositions and combinations comprising the interfering RNA molecules of the technology may be used in topical applications, for example in the form of insecticides.

[0110] In aspects, at least a portion of the nucleic acid compound of the vehicle may be released in an environment having a pH of from 3 to 10, such as from 4 to 9, from 5 to 8, from 6 to 7, or any range including and / or in between any two of these values. In aspects, greater than or equal to 60 wt.% of the nucleic acid compound may be released, such as greater than or equal to 65 wt.%, greater than or equal to 70 wt.%, greater than or equal to 75 wt.%, or greater than or equal to 80 wt.%, based on the total weight of the nucleic acid compound loaded in the vehicle.[OHl] 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. Accordingly, the claimed subject matter is not to be restricted except in light of the attached claims and their equivalents.EXAMPLES

[0112] Various aspects will be further clarified by the following examples, which are in no way intended to limit this disclosure thereto.Example 1: Preparation of initial glass powder

[0113] Exemplary glasses described herein are silicate glasses which include a base composition comprising, in weight percent on the oxide basis, of the constituents listed in Table 1.Table 1

[0114] The glasses were made in a platinum crucible using a batch of raw materials listed in Table 1. Each crucible containing the batch of raw materials was placed in a preheated furnace and were melted at 1100-1500 °C for 16 h. The glasses were refined to produce molten precursor glass that was then cast into a 7 inch x 11 inch x 0.5 inch block. The precursor glass was annealed at temperatures ranging from 400-650 °C, as noted in the Table 1. The glasses were crushed using air jet and ball mill to form initial glass powders (precursor glass particles), denoted as Example 1-1 and Example 1-2.Example 2: Acidic phosphate treatment of initial glass powders

[0115] The initial glass powders of Example 1-1 and Example 1-2 were modified to form glass powders comprising modified glass particles by treating the initial glass powders in a 0.5 M acidic phosphate solution in water at 60 °C. The initial glass powder used, duration of the treatment, and acidic phosphate solution of each example of Example 2 are shown in Table 2.Table 2

[0116] Identity of the crystalline phases of crystal phase assemblages and / or crystal sizes of a crystalline phase of the glass powders of Examples (Ex.) 2-1 through 2-4 were determined by X-ray diffraction (XRD) analysis techniques known to those in the art, using such commercially available equipment as the Bruker D4 Endeavor diffractometer manufactured by Bruker Corporation, Massachusetts, United States. Spectra were typically acquired for 29 from 10 to 100 degrees. XRD analysis was performed on the glass powders of Ex. 2-1 through 2-4 and the results obtained are as shown in Table 2. FIG. 1 shows XRD patterns of Ex. 2-1 110, Ex. 2-2 120, Ex. 2-3 130, and Ex. 2-4 140. The formation of brushite (CaHPCh^FFO), as indicated by the peaks marked with circles, and hydroxyapatite (Caio(P04)e(OH)2), as indicatedby the peaks marked with triangles, indicate the presence of both brushite and apatite on the surface of the modified glass particles, indicating good biocompatibility of these modified glass particles. Quantitative phase analysis of the XRD patterns using the Rietveld method was used to determine the wt.% 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.Table 3

[0117] 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 Figure 2. FIG. 2A shows Ex. 1-1 (untreated), FIG. 2B shows Ex. 2-1, FIG. 2C shows Ex. 2-2, FIG. 2D shows Ex. 1-2 (untreated), FIG. 2E shows Ex. 2-3, and FIG. 2F shows Ex. 2-4. The average particle size of the examples are summarized in Table 4.Table 4

[0118] The surface charge of the initial glass powders of Example 1 and the glass powders comprising modified glass particles of Example 2 was examined by measuring the zeta potential (Malvern Zetasizer Nano ZS) of the samples. The samples for measuring the zeta potential were prepared by suspending the glass particles in ethanol. The measured zetapotential of the initial glass powders and the acidic phosphate-treated glass powders are shown in Table 5.Table 5

[0119] The surface area and pore width of Ex. 1-1, Ex. 2-1, and Ex. 2-2 were analyzed by BET. The average BET surface area and pore width are summarized in Table 6. As shown in the data of Table 6, treatment of the initial glass powders with an acidic phosphate solution increased the BET surface area and decreased the pore width.Table 6Example 3: Nucleic acid loading on glass powders

[0120] In Example 3, a nucleic acid was loaded on the initial glass powders of Example 1 and the modified glass particles of Example 2 to form bioactive glass carriers. Specifically, glyceraldehyde 3-phosphate dehydrogenase having 195 base pairs (GADPH) was loaded onto Ex. 1-1, Ex. 2-1, Ex. 2-2, Ex. 1-2, Ex. 2-3, and Ex. 2-4 to determine a DNA payload estimation and binding rate. The GADPH serves as an artificial DNA surrogate that mimics a typicalRNAi fragment length. The GADPH loaded glass powders of Example 3 were prepared by combining 5 mg of the glass powder of Example 1 or Example 2 (5 mg / 40 pL ethanol) with a GADPH solution (6,000 ng / 60 pL H2O), mixing well by vortexing, and then incubating at room temperature for 30 min in a microtube equipped with gentle shaker (300 rpm). The samples were centrifuged at 14,000 rpm for 5 min to obtain 10 pL supernatant samples for DNA quantification by Qubit HS dsDNA assay. The DNA payload and binding rate of Examples 3-1 through 3-6 are reported in Table 7. As shown in the data of Table 7, treatment of the initial glass powders with an acidic phosphate solution increased the amount of the GADPH loaded onto the glass powder comprising the modified glass particles.Table 7Example 4: Nucleic acid elution from the bioactive glass carriers

[0121] After forming the bioactive glass carriers in Example 3, the elution efficiency of Examples 3-1 through 3-4 was evaluated under varying conditions. First, the bioactive glass carriers were centrifuged to remove all the supernatant prior to quantification. 300 pL of a phosphate buffer was added to 10 mg of each of the bioactive glass carriers, according to Table 8 and Table 9. The solution was resuspended by vortex and incubated at room temperature for 30 min using a gentle shaker (300 rpm). The solution was centrifuged at 14,000 rpm for 5 min to remove all the supernatant for nucleic acid quantification by Qubit HS dsDNA assay (10 pL sample for assay). The elution efficiency is reported in Table 8 and Table 9 as a percentage of nucleic acid released, relative to the total amount of nucleic acid loaded in Example 3. As acontrol, the bioactive glass carriers were also added to water and NaCl, but nucleic acid release was not detected.Table 8Table 9

[0122] The results of the nucleic acid loading in Example 4 and nucleic acid elution in Example 5, demonstrate the potential of the glass powders for nucleic acid transfer and delivery. Particularly, it was observed that the treatment of the initial glass powder with a phosphate salt, as described herein, can enhance the capacity of nucleic acid loading efficiency under neutral conditions. Further, the captured nucleic acid can be released in the slightly acidic buffer system with 30 minutes or less eluting time.Example 5: Nuclease resistance of bioactive glass carriers

[0123] The nuclease resistance of the bioactive glass carriers of Example 3 were evaluated. Specifically, the nucleic acid-loaded glass powders of Example 3 were subjected to harsh DNase challenging conditions to simulate nuclease attack under certain biological conditions. First, the bioactive glass carriers were centrifuged to remove all the supernatant prior to quantification. 180 pL of H2O was added to 5 mg of the nucleic acid-loaded glass powders of Example 3, followed by the addition of 20pL of 10X TURBO DNase buffer and 2 pL of TURBO DNase (4 units, where 1 unit is the amount of enzyme needed to degrade 1 pg of DNA in 10 min at 37 °C). The resulting mixture was incubated at 37 °C for 30 min. After incubating the mixture, the mixture was heated at 75 °C for 10 minutes, cooled to room temperature, and centrifuged at 15,500 rpm for 10 min. The supernatant of each sample was removed. The samples were then rinsed with 200 pL H2O, centrifuged at 15,500 rpm for 10 min, and all the supernatant was removed. The recovered DNA were examined using quantity determination by qPCR. The protecting rate was calculated by qPCR and is summarized in Table 10.Table 10

[0124] The recovered DNA were also examined using integrity confirmation by electrophoresis. The DNA quantity and integrity are compared with its corresponding DNase treated and non-treated sample. The electrophoresis image is shown in FIG. 3. The DNA input 302, DNA control 304, ladder 306, Ex. 3-3 308, Ex. 5-3 310, Ex 3-4 312, Ex. 5-4 314, ladder 316, Ex. 3-1 318, Ex. 5-1 320, Ex. 3-2 322, and Ex. 5-2 324 are shown in FIG 3. The relevant fragment of DNA at 195 bp is boxed and labeled in FIG. 3.Example 6: Cell internalization of bioactive glass carriers

[0125] Cell uptake efficiency and biocompatibility of the glass powders of Ex. 1-2 (untreated), Ex. 2-3 (KH2PO4 treated), and Ex. 2-4 (NaEEPCUP treated) were evaluated through in vitro testing. Specifically, Ex. 1-2, Ex. 2-3, and Ex. 2-4 were loaded with fluorescently labeled dsRNA to form the bioactive glass carriers. The fluorescent labeled dsRNA were observed inside the cultured insect cell sf9 after 48-hour co-culture with the bioactive glass carriers. As a control, dsRNA was cultured with the cells without a glass powder. FIG. 4 shows the fluorescent images and merged confocal images demonstrating the internalization of the bioactive glass carriers in the cells. The internalization of dsRNA and the bioactive glass carriers can be observed under fluorescent microscopy whereas no dsRNA signal was detected inside the cells using dsRNA alone. As the glass powders that are treated with an acidic phosphate solution have a higher nucleic acid payload, it is expected that a higher quantity of dsRNA is carried into the target cells relative to the non-treated example.Example 7: Efficacy of BAG-dsRNA particles in Colorado potato beetles

[0126] The effectiveness of BAG-dsRNA particles in silencing target genes and killing Colorado potato beetle (CPB) larvae was evaluated. Specifically, the BAG of Example 1-2 was loaded with the dsRNA targeting inhibitor of apoptosis, IAP gene, which was selected based on its known efficacy against CPB. The BAG-dsIAP particles were directly applied to leaf discs on a daily basis. These leaf discs were then fed to CPB larvae once a day for three consecutive days with 0.4pg dsIAP per larvae (10 larvae total) for statistical analysis. For comparison, a negative control was prepared in a similar manner in which the BAG of Example 1-2 was loaded with the dsGFP (green fluorescent protein).

[0127] The results are shown in FIG. 5. In particular, the mortality caused by BAG-dsIAP particles achieved 100% at day 5, which was comparable to the positive control (naked dsIAP). In addition, there was almost no mortality (90% survival rate) for the negative control with BAG-dsGFP, which demonstrated the low toxicity of the underlying BAG particles. The results demonstrate that the BAGs disclosed herein serve as RNA carriers with the ability to deliver such RNA to a cellular environment, such as in a leaf, which RNA remains efficacious in the leaf once consumed by CPB larvae.

[0128] Unless otherwise expressly stated, it is in no way intended that any method set forth herein be construed as requiring that its steps be performed in a specific order, nor that with any apparatus specific orientations be required. Accordingly, where a method claim does not actually recite an order to be followed by its steps, or that any apparatus claim does not actually recite an order or orientation to individual components, or it is not otherwise specifically stated in the claims or description that the steps are to be limited to a specific order, or that a specific order or orientation to components of an apparatus is not recited, it is in no way intended that an order or orientation be inferred, in any respect. This holds for any possible non-express basis for interpretation, including: matters of logic with respect to arrangement of steps, operational flow, order of components, or orientation of components; plain meaning derived from grammatical organization or punctuation, and; the number or type of aspects described in the specification.

[0129] All disclosed ranges are to be understood to encompass and provide support for claims that recite any and all subranges or any and all individual values subsumed by each range. For example, a stated range of 1 to 10 should be considered to include and provide support forclaims that recite any and all subranges or individual values that are between and / or inclusive of the minimum value of 1 and the maximum value of 10; that is, all subranges beginning with a minimum value of 1 or more and ending with a maximum value of 10 or less (e.g., 5.5 to 10, 2.34 to 3.56, and so forth) or any values from 1 to 10 (e.g., 3, 5.8, 9.9994, and so forth). Any listed range may be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a nonlimiting example, each range discussed herein may be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which may be subsequently broken down into subranges as discussed above. Finally, as will be understood by one 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 forth.

[0130] The drawings shall be interpreted as illustrating one or more aspects that are drawn to scale and / or one or more aspects that are not drawn to scale. This means the drawings may be interpreted, for example, as showing: (a) everything drawn to scale, (b) nothing drawn to scale, or (c) one or more features drawn to scale and one or more features not drawn to scale. Accordingly, the drawings can serve to provide support to recite the sizes, proportions, and / or other dimensions of any of the illustrated features either alone or relative to each other. Furthermore, all such sizes, proportions, and / or other dimensions are to be understood as being variable from 0-100% in either direction and thus provide support for claims that recite such values or any and all ranges or subranges that may be formed by such values.

[0131] The terms recited in the claims should be given their ordinary and customary meaning as determined by reference to relevant entries in widely used general dictionaries and / or relevant technical dictionaries, commonly understood meanings by those in the art, etc., with the understanding that the broadest meaning imparted by any one or combination of these sources should be given to the claim terms (e.g., two or more relevant dictionary entries should be combined to provide the broadest meaning of the combination of entries, etc.) subject only to the following exceptions: (a) if a term is used in a manner that is more expansive than its ordinary and customary meaning, the term should be given its ordinary and customary meaning plus the additional expansive meaning, or (b) if a term has been explicitly defined to have adifferent meaning by reciting the term followed by the phrase “as used in this document 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.). References to specific examples, use of “i.e.,” use of the word “technology,” etc., are not meant to invoke exception (b) or otherwise restrict the scope of the recited claim terms. Other than situations where exception (b) applies, nothing contained in this document should be considered a disclaimer or disavowal of claim scope.

[0132] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this technology belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the present application and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein. While not explicitly defined below, such terms should be interpreted according to their common meaning.

[0133] 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.

[0134] Unless the context indicates otherwise, it is specifically intended that the various features of the technology described herein may be used in any combination. Moreover, the disclosure also contemplates that in some aspects, any feature or combination of features set forth herein may be excluded or omitted. To illustrate, if the specification states that a complex comprises components A, B and C, it is specifically intended that any of A, B or C, or a combination thereof, may be omitted and disclaimed singularly or in any combination.

[0135] Unless explicitly indicated otherwise, all specified aspects, features, and terms intend to include both the recited aspect, feature, or term and biological equivalents thereof.

[0136] All patents, patent applications, provisional applications, and publications referred to or cited herein are incorporated by reference in their entirety, including all figures and tables, to the extent they are not inconsistent with the explicit teachings of this specification.

Claims

What is claimed is:

1. A glass powder comprising a plurality of modified glass particles, the plurality of modified glass particles comprising, in weight percent (wt.%): greater than or equal to 20 weight percent (wt.%) to less than or equal to 75 wt.% SiCh; 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 surface of the plurality of modified glass particles comprises a total amount of brushite and apatite of 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 upon contact.

2. The glass powder of claim 1, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 5 wt.% B2O3.

3. The glass powder of claim 1 or claim 2, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 5 wt.% AI2O3.

4. The glass powder of any preceding claim, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 30 wt.% Na2O.

5. The glass powder of any preceding claim, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 10 wt.% K2O.

6. The glass powder of any preceding claim, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 20 wt.% MgO.

7. The glass powder of any preceding claim, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 10 wt.% ZrCh.

8. The glass powder of any preceding claim, wherein the plurality of modified glass particles further comprise greater than 0 wt.% to less than or equal to 5 wt.% F.

9. The glass powder of any preceding claim, wherein the surface of the plurality of modified glass particles comprises an amount of brushite of greater than or equal to 10 wt.%of the total weight of the modified glass particles and an amount of apatite of greater than or equal to 15 wt.% of the total weight of the modified glass particles.

10. The glass powder of any preceding claim, wherein the surface of the plurality of modified glass particles comprises an amount of brushite of greater than or equal to 50 wt.% of the total weight of the modified glass particles and an amount of apatite of greater than or equal to 3 wt.% of the total weight of the modified glass particles.

11. The glass powder of any preceding claim, wherein a surface of the plurality of modified glass particles comprises a total amount of brushite and apatite of greater than or equal to 60 wt.% of the total weight of the modified glass particles.

12. The glass powder of any preceding claim, wherein the plurality of modified glass particles comprise a BET surface area of greater than or equal to 100 m2 / g to less than or equal to 300 m2 / g.

13. The glass powder of any preceding claim, wherein the plurality of modified glass particles comprise 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. The glass powder of any preceding claim, wherein the plurality of modified glass particles comprise a zeta-potential of greater than or equal to -10 mV to less than or equal to 5 mV.

15. The glass powder of any preceding claim, wherein the surface of the plurality of modified glass particles comprise acicular crystals having a largest length of greater than or equal to 10 nm to less than or equal to 50 nm.

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

17. The glass powder of any preceding claim, wherein the plurality of modified glass particles comprise: greater than or equal to 25 wt.% to less than or equal to 50 wt.% SiCE; 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.

18. The glass powder of any preceding claim, wherein the plurality of modified glass particles comprise: greater than or equal to 25 wt.% to less than or equal to 35 wt.% SiCh; 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.% ZrCh.

19. The glass powder of any preceding claim, wherein the plurality of modified glass particles comprise: greater than or equal to 38 wt.% to less than or equal to 48 wt.% SiCh; 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.

20. The glass powder of any preceding claim, wherein the plurality of modified glass particles further comprise the nucleic acid compound.

21. The glass powder of claim 20, 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 combinations thereof.

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

23. A method of forming a glass powder, the method comprising: processing a glass into precursor glass particles, wherein the precursor glass particles have an average longest dimension of greater than or equal to 0.5 micrometers (pm) to less than or equal to 10 pm; and treating the precursor glass particles with a treatment solution, wherein: the treatment solution comprises a phosphate salt; and the treating forms modified glass particles; wherein:a surface of the plurality of modified glass particles comprises a total amount of brushite and apatite of greater than or equal to 30 wt.% of the total weight of the modified glass particles; and the plurality of modified glass particles can bind to a nucleic acid compound upon contact.

24. The method of claim 23, wherein a pH of the treatment solution is of greater than or equal to 2.5 to less than or equal to 4.5.

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

26. The method of any one of claims 23 to 25, wherein the treatment solution comprises sodium phosphate, potassium phosphate, or a combination thereof.

27. The method of any one of claims 23 to 26, wherein the treating comprises: forming a mixture comprising the precursor glass particles and the treatment solution; and heating the mixture at a temperature of from 40 °C to 80 °C for a 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 a glass into precursor glass particles, wherein the precursor glass particles have an average longest dimension of greater than or equal to 0.5 micrometers (pm) to less than or equal to 10 pm; and treating the precursor glass particles with a treatment solution, wherein: the treatment solution comprises a phosphate salt; and the treating forms modified glass particles; contacting the plurality of modified glass particles with a nucleic acid compound to form an admixture; andincubating the admixture under conditions in which the nucleic acid compound non- covalently binds to the plurality of modified glass particles, forming the bioactive glass carrier.

30. The method of claim 29, wherein the admixture comprises a concentration of the nucleic acid compound 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 claim 30, wherein the incubating the admixture is conducted at a temperature of greater than or equal to 20 °C to less than or equal to 40 °C.

32. The method of claim any one of claims 29 to 31, wherein the incubating the admixture is conducted for a period greater than or equal to 10 minutes.

33. The method of any one of claims 29 to 32, wherein the incubating the admixture comprises loading the nucleic acid compound on the plurality of modified glass particles at a loading efficiency of greater than or equal to 50%.

34. The method of any one of claims 29 to 33, wherein the bioactive glass carrier comprises a concentration of the nucleic acid compound 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 vehicle of claim 35, wherein the vehicle comprises a drug delivery vehicle, a gene delivery vehicle, a tissue regeneration vehicle, a crop treatment or crop protection vehicle, an immunomodulatory vehicle, or a gene editing complex vehicle.

37. The vehicle of claim 35 or claim 36, wherein at least a portion of the nucleic acid compound is released at a pH greater than or equal to 3 to less than or equal to 10.

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

39. A method of releasing the nucleic acid compound from the glass powder of any one of claims 20-22 or 35-38, the method comprising:positioning the glass powder in a cellular environment, wherein at least a portion of the nucleic acid compounds 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 greater than or equal to 3 to less than or equal to 10.

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

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

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