BORATE-GLASS BIOMATERIALS

Abstract

Borate-glass biomaterials comprising: aNa.sub.2O. bCaO. cP.sub.2O.sub.5. dB.sub.2O.sub.3 wherein a is from about 1-40 wt %, b is from about 10-40 wt %, c is from about 1-40 wt %, and d is from about 35-80 wt %; and wherein the biomaterial has a surface area per mass of more than about 5 m.sup.2/g. Methods of making and uses of these biomaterials.

Claims

1-44. (canceled)

45. A borate-glass biomaterial comprising a composition having a B.sub.2O.sub.3 component, a CaO component, and at least one other component selected from a P.sub.2O.sub.5 component and a Na.sub.2O component, wherein the B.sub.2O.sub.3 component is a main network forming component and the biomaterial has a surface area per mass of more than about 5 m.sup.2/g.

46. The borate-glass biomaterial of claim 45, wherein the composition comprises: aNa.sub.2O. bCaO. cP.sub.2O.sub.5. dB.sub.2O.sub.3, wherein a is from about 1-40 wt %, b is from about 10-40 wt %, c is from about 1-40 wt %, and d is from about 35-80 wt %.

47. The borate-glass biomaterial of claim 46, wherein a is from about 15-30 wt %, b is from about 15-30 wt %, c is from about 3-7 wt %, and d is from about 35-65 wt %.

48. The borate-glass biomaterial of claim 45, wherein the composition comprises: xCaO. yP.sub.2O.sub.5. zB.sub.2O.sub.3, wherein x is from about 5-50 wt %, y is from about 5-50 wt %, and z is from about 35-75 wt %, or x is 10-50 wt %, y is 5-35 wt % and z is 38-80 wt %.

49. The borate-glass biomaterial of claim 45, wherein the composition comprises: 1Na.sub.2O. mCaO nB.sub.2O.sub.3. wherein 1 is from about 5-50 wt %, m is from about 1-50 wt %, and n is from about 40-80 wt %.

50. The borate-glass biomaterial of claim 45, wherein the biomaterial has a surface area per mass of about 5-300 m.sup.2/g, 10-300 m.sup.2/g, 20-300 m.sup.2/g, 30-300 m.sup.2/g, 40-300 m.sup.2/g, 50-300 m.sup.2/g, 60-300 m.sup.2/g, 70-300 m.sup.2/g, 80-300 m.sup.2/g, 90-300 m.sup.2/g, 100-300 m.sup.2/g, 110-300 m.sup.2/g, 120-300 m.sup.2/g, 130-300 m.sup.2/g, 140-300 m.sup.2/g, 150-300 m.sup.2/g, 200-300 m.sup.2/g, 250-300 m.sup.2/g, 5-250 m.sup.2/g, 5-200 m.sup.2/g, 5-150 m.sup.2/g or 5-100 m.sup.2/g.

51. The borate-glass biomaterial of claim 45, wherein the biomaterial has a pore volume per mass of biomaterial of about 0.1-3.0 cm.sup.3/g, 0.2-3.0 cm.sup.3/g, 0.3-3.0 cm.sup.3/g, 0.4-3.0 cm.sup.3/g, 0.5-3.0 cm.sup.3/g, 0.6-3.0 cm.sup.3/g, 0.7-3.0 cm.sup.3/g, 0.8-3.0 cm.sup.3/g, 0.9-3.0 cm.sup.3/g, 1.0-3.0 cm.sup.3/g, 0.1-2.5 cm.sup.3/g, 0.42-1.18 cm.sup.3/g or 0.1-2.0 cm.sup.3/g.

52. The borate-glass biomaterial of claim 45, wherein the biomaterial can induce bone formation.

53. The borate-glass biomaterial of claim 45, wherein the biomaterial is one of amorphous, crystalline or semi-crystalline.

54. The borate-glass biomaterial of claim 45, wherein the biomaterial comprises at least one of: particles, fibrils, hollow spheres, solid spheres, monoliths, fibrous form or a porous sponge scaffold.

55. The borate-glass biomaterial of claim 45, wherein the biomaterial comprises particles having a diameter of 0.2-1 μm, 5-2000 μm, 5-100 μm, or 25-75 μm.

56. The borate-glass biomaterial of claim 45, further comprising a carrier.

57. The borate-glass biomaterial of claim 45, wherein the biomaterial is coated on a bone implant surface.

58. A method for making the borate-glass biomaterial of claim 45, comprising combining a boron precursor and a calcium precursor, with at least one of a phosphate precursor and a sodium precursor to form a mixture; gelling the mixture to form a gel; drying the gel; and calcining the dried gel, wherein the boron precursor solution is selected from trimethyl borate B(OCH.sub.3).sub.3, triethyl borate B(C.sub.2H.sub.5O).sub.3, tributyl borate B(CH.sub.3(CH.sub.2).sub.3O).sub.3, Tri-tert-butyl borate (B.sub.3(CH.sub.3).sub.3CO) and boric acid, and wherein the calcium precursor is selected from calcium methoxyethoxide, Calcium nitrate tetrahydrate (Ca(NO.sub.3).sub.24H.sub.2O), Calcium Chloride (CaCl.sub.2), Calcium Ethoxide (Ca(C.sub.2H.sub.5O).sub.2), and Calcium methoxide (C.sub.2H.sub.6CaO.sub.2).

59. The method of claim 58, wherein the phosphate precursor is selected from triethyl phosphate, Trimethyl phosphate ((CH.sub.3).sub.3PO.sub.4), Tributyl phosphate ((CH.sub.3CH.sub.2CH.sub.2CH.sub.2O).sub.3PO), Dibutyl phosphate ((CH.sub.3CH.sub.2CH.sub.2CH.sub.2O).sub.2P(O)OH), n-Butyl phosphate, mixture of monobutyl and dibutyl (C.sub.8H.sub.19O.sub.4P/C.sub.4H.sub.11O.sub.4P).

60. The method of claim 58, wherein the sodium precursor is selected from Sodium methoxide (NaCH.sub.3O) in methanol (different %) and sodium hydroxide (NaOH).

61. The method of claim 58, wherein the mixture comprises boric acid, anhydrous ethanol, triethyl phosphate, calcium methoxyethoxide, and sodium methoxide.

62. The method of claim 58, wherein gelling the mixture comprises maintaining the solution at a temperature between about room temperature and about 60° C., preferably at about 37° C.

63. The method of claim 58, wherein calcining the dry gel comprises heating the dry gel to between about 400-600° C., or about 100-400° C.

64. The borate-glass biomaterial of claim 45, for use in at least one of: mineralization, reducing dentine sensitivity, bone regeneration, wound healing, filling hard or soft tissue defects, as a coating on a bone implant, for enhancing the appearance of skin, or as a drug delivery vehicle.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0070] Further aspects and advantages of the present invention will become better understood with reference to the description in association with the following in which:

[0071] FIG. 1 is a schematic of an embodiment of a method for making a biomaterial according to the present disclosure.

[0072] FIG. 2 shows x-ray diffraction (XRD) spectra of the biomaterials of Example 1, according to embodiments of the present disclosure.

[0073] FIG. 3 shows ATR-FTIR spectra of the biomaterials of Example 1 calcined at different temperatures, according to embodiments of the present disclosure.

[0074] FIG. 4 shows NMR spectra of the biomaterials of Example 1, according to embodiments of the present disclosure.

[0075] FIG. 5 shows relationship between crystallization temperature (T.sub.c), glass transition temperature (T.sub.g)and N.sub.C for the biomaterials of Example 1, according to embodiments of the present disclosure.

[0076] FIG. 6 shows Dynamic Vapour Sorption (DVS) for the biomaterials of Example 1, according to embodiments of the present disclosure.

[0077] FIG. 7 shows release of (a) boron, (b) calcium, (c) sodium and (d) phosphorus ions from the biomaterials of Example 1, according to embodiments of the present disclosure, and the comparative biomaterial of Example 2, as a function of time in distilled water.

[0078] FIG. 8 shows ATR-FTIR spectra of the biomaterials of Example 1 at different time points after immersion in simulated body fluids (SBF), according to embodiments of the present disclosure.

[0079] FIG. 9 shows XRD spectra of the biomaterials of Example 1 at different time points after immersion in simulated body fluids (SBF), according to embodiments of the present disclosure.

[0080] FIG. 10 shows the change in pH over dissolution time in SBF (solid line) and deionized water (DIW) (dashed line) of the biomaterials of Example 1, according to different embodiments of the present disclosure, and the comparative biomaterial of Example 2.

[0081] FIG. 11 shows scanning electron micrographs of the surface of (a) one of the biomaterials of Example 1 (B46) according to an embodiment of the present disclosure, and (b) the comparative biomaterial of Example 2 (45B5), after calcination, and after 6 hours and 7 days in simulated body fluids.

[0082] FIG. 12 shows the change in pH over dissolution time in SBF, K.sub.2HPO.sub.4 and deionized water (DIW) of one of the biomaterials of Example 1 (B46), according to an embodiment of the present disclosure, and the comparative biomaterial of Example 2 (45B5).

[0083] FIG. 13 are XRD spectra of one of the biomaterials of Example 1 (B46), according to an embodiment of the present disclosure, and the comparative biomaterial of Example 2 (45B5) at different time points in SBF and K.sub.2HPO.sub.4 solution.

[0084] FIG. 14 are ATR-FTIR spectra of one of the biomaterials of Example 1 (B46), according to an embodiment of the present disclosure, and the comparative biomaterial of Example 2 (45B5) at different time points in SBF and K.sub.2HPO.sub.4 solution.

[0085] FIG. 15 is a graph showing the effect of calcination temperature on one of the biomaterials (B46) of Example 1, according to embodiments of the present disclosure, on surface area and pore volume.

[0086] FIG. 16 (a) and (b) show % mesenchymal stem cell viability in the presence of ionic release from the biomaterials of Example 1, according to embodiments of the present disclosure, at dilutions of 1:1 and 1:4 respectively.

DETAILED DESCRIPTION OF THE INVENTION

[0087] This invention is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the drawings. The invention is capable of other embodiments and of being practiced or of being carried out in various ways. Also, the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. The use of “including”, “comprising”, or “having”, “containing”, “involving” and variations thereof herein, is meant to encompass the items listed thereafter as well as, optionally, additional items. In the following description, the same numerical references refer to similar elements. In the drawings, like reference characters designate like or similar parts.

[0088] One aspect of the present disclosure is directed to biomaterials which comprise a borate-glass. In one embodiment, the biomaterial comprises: aNa.sub.2O. bCaO. cP.sub.2O.sub.5. dB.sub.2O.sub.3, wherein a is from about 1-40 wt %, b is from about 10-40 wt %, c is from about 1-40 wt %, and d is from about 35-80 wt %; and wherein the biomaterial has a surface area per mass of more than about 5 m.sup.2/g.

[0089] In another embodiment, the biomaterial comprises: xCaO. yP.sub.2O.sub.5. zB.sub.2O.sub.3, wherein x is from about 5-50wt %, y is from about 5-50 wt %, and z is from about 35-75 wt %; and wherein the biomaterial has a surface area per mass of more than about 5 m.sup.2/g.

[0090] In another embodiment, the biomaterial comprises: INa.sub.2O. mCaO. nB.sub.2O.sub.3, wherein 1 is from about 5-50 wt %, m is from about 1-50 wt %, and n is from about 40-80 wt %; and wherein the biomaterial has a surface area per mass of more than about 5 m.sup.2/g. In certain embodiments of any of the foregoing or following, the biomaterial has a surface area per mass of more than about 10 m.sup.2/g, more than about 20 m.sup.2/g, more than about 30 m.sup.2/g, more than about 40 m.sup.2/g, or more than about 50 m.sup.2/g.

[0091] The surface area per mass of the biomaterial is about 5-300 m.sup.2/g, 10-300 m.sup.2/g, 20-300 m.sup.2/g, 30-300 m.sup.2/g, 40-300 m.sup.2/g, 50-300 m.sup.2/g, 60-300 m.sup.2/g, 70-300 m.sup.2/g, 80-300 m.sup.2/g, 90-300 m.sup.2/g, 100-300 m.sup.2/g, 110-300 m.sup.2/g, 120-300 m.sup.2/g, 130-300 m.sup.2/g, 140-300 m.sup.2/g, 150-300 m.sup.2/g, 200-300 m.sup.2/g, 250-300 m.sup.2/g, 5-250 m.sup.2/g, 5-200 m.sup.2/g, 5-150 m.sup.2/g or 5-100 m.sup.2/g. The pore volume per mass of biomaterial is about 0.1-3.0 cm.sup.3/g, 0.2-3.0 cm.sup.3/g, 0.3-3.0 cm.sup.3/g, 0.4-3.0 cm.sup.3/g, 0.5-3.0 cm.sup.3/g, 0.6-3.0 cm.sup.3/g, 0.7-3.0 cm.sup.3/g, 0.8-3.0 cm.sup.3/g, 0.9-3.0 cm.sup.3/g, 1.0-3.0 cm.sup.3/g, 0.1-2.5 cm.sup.3/g, 0.42-1.18 cm.sup.3/g or 0.1-2.0 cm.sup.3/g.

[0092] In one embodiment the biomaterial is amorphous and is in particulate form with a particle size of about 5-2000 μm. The biomaterial may be used for bone regeneration or augmentation and may be placed in a bone defect or at a site requiring bone augmentation.

[0093] The biomaterial may be made into a slurry using the patient's own blood or bone before packing into the defect. The biomaterial may also be injected. From another aspect, there is provided a method 100 of making the biomaterials described herein (FIG. 1).

[0094] In one embodiment of the method 100, the method comprises an adapted sol-gel method. Briefly, the method comprises forming a solution to be gelled 102, gelling the solution to form a gel 104, drying the gel to form a dry gel 106, calcining the dry gel to remove organic matter 108, and optionally grinding the calcined dry gel, and/or sizing the calcined dry gel to obtain particles within a certain size and/or shape range.

[0095] In one embodiment, forming the solution comprises mixing together precursors. The precursors comprise boric acid, triethyl phosphate, calcium methoxyethoxide (20% in methoxyethanol), and sodium methoxide. Gelling the solution comprises casting the solution into sealed vials and placing in an oven at 37° C. The gelled solution may also be left to age at room temperature or elevated temperatures for example for about 2-15 days. In one embodiment, drying the gels comprises removing from the oven and placing in crystallization dishes to dry at room temperature for a day then in an oven at 120° C. for 2 days. In some embodiments, the drying step causes the gel to dry to a particulate form. Calcining the dry gel comprises heating (400-600° C.) in air using a 3° C./min heating rate, 2 hour dwell, and furnace cooling. In certain embodiments, the method comprises grinding the calcined dry gel and sieving to a desired size range such as 25-75 μm.

[0096] Other starting solutions (precursors) can include: [0097] For Boron: Trimethyl borate B(OCH.sub.3).sub.3, triethyl borate B(C.sub.2H.sub.5O).sub.3, tributyl borate B(CH.sub.3(CH.sub.2).sub.3O).sub.3, Tri-tert-butyl borate (B.sub.3(CH.sub.3).sub.3CO) [0098] For Calcium: Calcium nitrate tetrahydrate (Ca(NO.sub.3).sub.2.4H.sub.2O), Calcium Chloride (CaCl.sub.2), Calcium Ethoxide (Ca(C.sub.2H.sub.5O).sub.2), Calcium methoxide (C.sub.2H.sub.6CaO.sub.2) [0099] For Sodium: Sodium methoxide (NaCH.sub.3O) in methanol (different %), Sodium hydroxide (NaOH) [0100] For Phosphate: Trimethyl phosphate ((CH.sub.3).sub.3PO.sub.4), Tributyl phosphate ((CH.sub.3CH.sub.2CH.sub.2CH.sub.2O).sub.3PO), Dibutyl phosphate ((CH.sub.3CH.sub.2CH.sub.2CH.sub.2O).sub.2P(O)OH), n-Butyl phosphate, mixture of monobutyl and dibutyl (C.sub.8H.sub.19O.sub.4P/C.sub.4H.sub.11O.sub.4P)

[0101] A second embodiment of the method differs from the first embodiment in that sodium ions are not included. In this case, the method comprises mixing boric acid, triethyl phosphate and calcium methoxyethoxide (20% in methoxyethanol) and following the same procedure as described for the first embodiment.

[0102] A third embodiment of the method differs from the first embodiment in that phosphorus ions are not included. In this case, the method comprises mixing boric acid, sodium methoxide and calcium methoxyethoxide (20% in methoxyethanol) and following the same procedure as described for the first embodiment.

[0103] Identification of equivalent compositions and methods are well within the skill of the ordinary practitioner and would require no more than routine experimentation, in light of the teachings of the present disclosure. Practice of the disclosure will be still more fully understood from the following examples, which are presented herein for illustration only and should not be construed as limiting the disclosure in any way.

EXAMPLES

Example 1

Method of Making Borate-Glass Biomaterials

[0104] Borate-glass biomaterials, according to certain embodiments of the present disclosure, were made using a sol-gel process. All sol-gel processing took place within a nitrogen purged glove box. Four component borate glass compositions based on boron, Ca, Na and P were made by incrementally increasing or decreasing the amount of boron while maintaining the ratio of the Ca, Na, and P (Table 1). Boric Acid (>99.5%) was first added to anhydrous ethanol in a Teflon beaker which was covered with a watch glass, stirred magnetically, and heated to about 35-50° C., preferably 40° C.±3° C., to aid dissolution. Advantageously, unlike most known sol gel methods, a mild heating of about 35-50° C. suffices. After the solution became clear, triethyl phosphate (>99.8%), calcium methoxyethoxide (20% in methoxyethanol), and sodium methoxide (25 wt. % in methanol) were added drop wise in 30 minute intervals. After the final addition the solution was mixed for another 30 minutes or until the viscosity became too great for stirring.

[0105] The sol was then cast into polypropylene vials (4 mL capacity. O. D.×Height: 13×57 mm), sealed, and stored in an oven at 37° C. for further gelation and ageing. In some instances, gelation, or partial gelation, occurred within the mixing vessel before casting. After 10 days the gels were removed as monolithic gels and placed in crystallization dishes and dried in air at room temperature for one day, then in an oven at 120° C. for 2 days. During the first part of the drying, all gel monoliths collapsed into particles. The collapsing of the monolithic gel is often observed with non-silicate sol-gel systems and is extremely difficult to avoid even with controlled methods. In order to prevent this collapse and to create a monolith structure, the gel can be combined with a polymer or binder and dried under controlled conditions such as those using critical point drying.

[0106] Monolithic gels also formed in larger containers (60 mL capacity:. O.D.×Height: 53×47 mm) demonstrating the ability to scale-up this process. The particles were then calcined in air to various temperatures (400° C., 450° C., 500° C., 550° C. and 600° C.) using a 3° C./min heating rate, 2 hour dwell time, and then furnace cooled. Lower calcination temperatures (100° C. to 400° C.) are also possible. Following calcination, the particles were ground and sieved to isolate a particle size fraction of 25-75 μm and stored in a desiccator until analysis. For traditional, melt derived bioactive glasses, sodium is often added to help reduce the firing temperature of the glass melt but for sol-gel derived glasses this is not necessary due to the low processing conditions. However to directly compare the biomaterial of the present disclosure to melt-derived biomaterials of similar compositions (Example 2), sodium was added.

TABLE-US-00001 TABLE 1 Biomaterial compositions made in Example 1 according to embodiments of the present disclosure. wt. % (mol %) ID B.sub.2O.sub.3 CaO Na.sub.2O P.sub.2O.sub.5 B36 38.6 (36.2) .sup. 27.4 (31.90 27.4 (28.9) 6.6 (3.0) B41 43.6 (41.1) 25.2 (29.4) 25.2 (26.6) 6.1 (2.8) B46 48.6 (46.1) 22.9 (26.9) 22.9 (24.4) 5.6 (2.6) B51 53.6 (51.1) 20.7 (24.4) 20.7 (22.1) 5.1 (2.4) B56 58.6 (56.1) 18.4 (21.9) 18.4 (19.8) 4.6 (2.2) B61 63.6 (61.3) 16.2 (19.3) 16.2 (17.5) 4.1 (1.9)

[0107] Each biomaterial formulation of Table 1 successfully underwent gelation even at low boron concentrations as confirmed qualitatively by holding the storage vials upside down and noting no flow at room temperature, although for the lowest boron concentration, B36, gelation did not occur until the second day.

[0108] The pH of the solution was measured immediately after addition of the sodium precursor and before casting, and were as follows: B36, pH 13.3; B41, pH 13.1; B46, pH 13.1; B51, pH 12.9; B56, pH 12.7 and B61, pH 11.2. During addition of the precursors, the pH was found to increase rapidly from about 3 to about 11 after addition of the calcium precursor.

[0109] Instead of boric acid being added to ethanol, traditional sol gel precursors of triethyl borate, trimethyl borate and tributyl borate were individually tried within the method of Example 1. However, with these precursors, there was no gel formation suggesting no network formation although they did form a glass.

Example 2

Comparative Example

[0110] To compare the effect of processing methods, a borate-glass biomaterial with a substantially equivalent composition (Table 2) to that of B46 of Example 1 was created using a melt quench technique. Boric Acid, monosodium phosphate, sodium carbonate, and calcium carbonate were thoroughly dry mixed and placed in a Pt crucible then heated at 1100° C. for 2 hours with intermediate stirring to insure homogeneity. The melt was then rapidly quenched between two steel plates and the resultant glass ground to 25-75 μm particles. The comparative biomaterial thus obtained, and the method used to obtain it, does not form part of the present disclosure and is included here for comparison purposes only.

TABLE-US-00002 TABLE 2 Comparative example of melt-derived composition. wt. % (mol %) ID B.sub.2O.sub.3 CaO Na.sub.2O P.sub.2O.sub.5 Comparative 48.6 (46.1) 22.9 (26.9) 22.9 (24.4) 5.6 (2.6) Example 45B5

Example 3

Effect of Calcination Temperature on Crystallinity of the Glasses of Example 1

[0111] The effect of calcination temperature (400-600° C., and as-made (AM)) on structure for the sol-gel glasses of Example 1 was investigated using X-ray diffractometry (XRD). The sol-gel derived glass powders of Example 1 were analyzed with a Bruker D8 Discover™ X-ray diffractometer equipped with a CuKα (λ=0.15406 nm) target set to a power level of 40 mV and 40 mA. Using an area detector, three frames of 25° were collected from 15-75 2 theta) (°) and merged in post processing. Phase identification was carried out using X'Pert Highscore Plus™.

[0112] FIG. 2 illustrates the x-ray diffraction spectra for the compositions of Table 1. All the biomaterials of Example 1 were amorphous at a 400° C. calcination temperature which suggests homogeneity. Progressive crystallization was observed with increasing calcination temperature, with low borate content glasses crystallizing at lower temperatures. The lower boron containing formulations began to crystallize before 450° C. The as-made glass particles displayed two broad humps indicating their amorphous and homogenous nature except for B36 which displayed a number of minor peaks, attributable to a precipitate phase. At 600° C. all Example 1 compositions were crystallized into various sodium calcium borates verified by XRD and FTIR, indicating the successful incorporation of the precursors into the glass network during the sol-gel processing (FIG. 3). It was also found that the compositions with higher sodium content tended to crystallize at lower temperatures, which was corroborated by ATR-FTIR spectroscopy which showed sharp, doublet peaks at increased calcination temperatures, indicating crystallization. Since all Example 1 compositions remained amorphous post calcinations at 400° C., and amorphous materials tend to be more bioactive, compositions calcined at 400° C. were chosen to compare the properties of the various Example 1 glasses.

Example 4

Thermal Analysis

[0113] Differential scanning calorimetry (DSC) (Jupiter STA 449™) was performed using 30 mg of calcined glass powder in a Pt crucible under flowing argon purge. Analysis was carried out between 50 and 1000° C. at a heating rate of 10° C./min, followed by furnace cooling to room temperature. The output was used to calculate glass transition and crystallization temperatures (T.sub.g and T.sub.c, respectively), which are presented in Table 3.

TABLE-US-00003 TABLE 3 Glass transition temperature (T.sub.g) and crystallization temperature (T.sub.c) of embodiments of biomaterials according to present disclosure (B36, B41, B46, B51, B56 and B61) and comparative example 45B5 ID T.sub.g (° C.) T.sub.c (° C.) B36 431 474 B41 441 510 B46 453 525 B51 474 579 B56 485 632 B61 484 639 Comparative Example 45B5 473 531

[0114] An increase in both T.sub.g and T.sub.c was observed with increasing in glass borate content. The DSC also corroborated the crystallization behavior observed through XRD in Example 3.

Example 5

Textural Particle Properties

[0115] The sieved particle sizes and porosities of the sol-gel glass compositions of Example 1 (all calcined at 400° C.) were compared to those of the comparative melt-derived glass particles of Example 2. Particle sizes (D.sub.50) were determined using a sedigraph (Horiba LA-920™). The specific surface areas of the calcined powders (400° C., n=3) were measured with N.sub.2 (g) adsorption and desorption isotherms collected with a Micrometics TriStar 3000™ (Micromeritics Instrument Corporation, Norcross, Ga.) gas sorption system. The specific surface areas were determined from the isotherm with the BrunauerEmmettTeller (BET) method (S. Brunauer, P. H. Emmett, E. Teller, Adsorption of gases in multimolecular layers. Journal of the American Chemical Society 60, 309-319 (1938)). The average pore width and pore volume was provided using the adsorption isotherms using the Barrett-Joyner-Halenda (BJH) method (L. G. Joyner, E. P. Barrett, R. Skold, The Determination of Pore Volume and Area Distributions in Porous Substances. II. Comparison between Nitrogen Isotherm and Mercury Porosimeter Methods. Journal of the American Chemical Society 73, 3155-3158 (1951); published online Epub1951/07/01 (10.1021/ja01151a046)). The results are presented in Table 4.

TABLE-US-00004 TABLE 4 Textural particle properties of embodiments of biomaterials according to present disclosure (B36, B41, B46, B51, B56 and B61) and comparative example 45B5 Pore D50 SSA Width Pore Volume ID (μm) (m.sup.2/g) (nm) (cm.sup.3/g) B36 34.7 54.9 ± 7.7 32.8 ± 2.0 0.42 ± 0.06 B41 34.6 71.8 ± 8.3 33.2 ± 1.9 0.65 ± 0.11 B46 43.8 93.8 ± 8.2 28.9 ± 0.7 0.74 ± 0.05 B51 33.7 114.2 ± 14.9 32.9 ± 1.0 0.94 ± 0.18 B56 38.8 121.0 ± 12.9 29.6 ± 0.4 0.98 ± 0.11 B61 47.1 138.4 ± 11.8 29.0 ± 0.7 1.18 ± 0.12 Comparative 44.1  0.238 ± 0.017 34.0 ± 8.6 0.00089 ± 0.00006 Example 45B5

[0116] As can be seen, embodiments of the biomaterials of the present dislcosure made using an embodiment of the sol-gel method of the present disclosure have specific surface area and total pore volume values much higher than the melt-derived comparative example, ˜400 and ˜800 times more respectively.

[0117] Furthermore, the specific surface area and total pore volume could be controlled by varying the boron concentration. SSA and pore volume increased with increasing glass borate content. Reduction in surface area is the driving force in densification and the biomaterials with the lowest boron concentrations have the lowest specific surface area and total pore volume suggesting they are the most dense. This is supported by XRD which shows that the low boron content glasses crystallize at the lowest temperatures (FIG. 2). Furthermore, this trend is not due particle size because the glass with the highest surface area also has the largest particle diameter (D.sub.50). In contrast, the average pore width remained consistent which may be attributable to uniform processing conditions. An increase in the calcination temperature of B46 (calcination at 400° C., 450° C., 500° C., 550° C. and 600° C.) of Example 1, led to a decrease in SSA and pore volume.

Example 6

Structural Properties

[0118] Each biomaterial of Example 1 remained amorphous according to XRD and the structure was further examined by Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) and Solid state .sup.11B nuclear magnetic resonance (NMR). For the ATR-FTIR, a Spectrum 400™ (Perkin-Elmer) was used to collect spectra in a wavenumber range between 4000 and 650 cm.sup.−1 .sup.1with a resolution of 4 cm.sup.−1 using 64 scans per sample. All spectra were baseline corrected and normalized to the total area surface area under absorption bands using Spectrum software (Perkin-Elmer, USA). For the NMR, .sup.11B NMR spectra were recorded on an Agilent/Varian VNMRS300 spectrometer with a .sup.11B frequency of 96.2 MHz. Approximately 2000 accumulations for each spectra were obtained by applying a 90 degree pulse of microseconds every 8 seconds with proton decoupling.

[0119] The three main regions associated with borate-based glasses were present at 850-1200 cm.sup.−1 (B—O stretching of BO.sub.4.sup.−units), 1200-1500 cm.sup.−1 (B—O stretching of BO.sub.3 units), and a band at ˜720 cm.sup.−1 attributable to the B—O—B bending of BO.sub.3.sup.3− units, which was more defined in lower borate content glasses (FIG. 3). With a decrease in glass borate content, the broad band between ˜942 and ˜1000 cm.sup.−1, attributable to the B—O linkages of BO.sub.4.sup.−, formed a defined shoulder peak at ˜870 cm.sup.−1, which is characteristic of the B—O stretching of boroxoal rings as observed in the B36 to B51 range of the Example 1 glasses. A comparison of the spectra of 45B5 (Comparative Example 2) and B46 (Example 1) indicated that both glasses were of analogous structures as they displayed similar peaks. In FIG. 3 it can be seen that as the calcining temperature increases the glasses become more crystalline as seen by the sharp peaks on the XRD diffractograms. Lower borate containing glasses crystallize first likely due to their poorly connected glass network which allows for quicker densification.

[0120] .sup.11B MAS NMR spectroscopy (FIG. 4) provided geometrical information on borate unit of the biomaterials of Examples 1 and 2. All biomaterials exhibited a large, fairly sharp peak near 0 ppm which can be attributed to .sup.11B nuclei occupying a fairly symmetric site in chemical structure. The small broadening to the right of this peak, as seen in B46-B61, may be due to .sup.11B in less symmetric sites. B36, B41 and B61 showed greater broadening on the left side which may indicate .sup.11B in asymmetric sites (i.e. BO.sub.3) or the same amount but in sites of less symmetry. The resonance associated with B0.sub.4 was quite narrow located in a chemical shift range of 0 and −11 ppm due to its small quadrupole coupling constant. Conversely, BO.sub.3 had a stronger quadropole interaction and produced a more broad resonance between 14 and 18 ppm. Further, when .sup.11B occupies a state of low symmetry, the relaxation times are relatively short (100 ms) due to its quadrupolar nucleus. The glasses analyzed in this study required an 8 second delay between pulses suggesting the .sup.11B were in a state of high symmetry and likely tetrahedrally coordinated, supporting the ATR-FTIR spectra.

Example 7

Calculation of Glass Network Connectivity

[0121] Network connectivity (N.sub.C) is a measure of the bridging oxygen bonds per network former (usually calculated for an Si atom in silicate-based glasses) and has been used to predict the bioactivity of glasses. N.sub.C is measured on a scale of 0 to 4, with 4 indicating a fully connected, chemically most stable network (e.g., quartz). On the other hand, glasses with an N.sub.C between 2 and 2.6 have generally been regarded as bioactive (e.g. Edén M. Journal of Non-Crystalline Solids. 2011; 357:1595-602), e.g., Bioglass® (45S5), which has an N.sub.C of 2.12 (Hill R G et al Journal of Non-Crystalline Solids. 2011; 357:3884-7) as calculated using Equation (1).

[00001]$\begin{array}{cc}{N}_C=\frac{4.Math.\left({\mathrm{SiO}}_2\right)-2\left[M_2^I.Math.O+M^{\mathrm{II}}.Math.O\right]+6\left[P_2.Math.O_5\right]}{\left[{\mathrm{SiO}}_2\right]}& \left(1\right)\end{array}$

[0122] where M.sup.1 and M.sup.11 represent glass network modifiers sodium and calcium, respectively. Modeling and NMR studies, have indicated that the phosphorous does not enter the glass network (i.e., no Si—O—P bonds are formed) and remains as an orthophosphate [PO.sub.4.sup.3−], which is accounted for in the above calculation. However, Si—O—P bonds can occur at higher P concentrations (>50 mol %) [8].

[0123] In the case of borate-based glasses, while it is also possible to form B—O—P bonds, as in the case of silicate-based glasses, it is assumed that the phosphorous does not enter the glass network and is present in an orthophosphate. In addition, if it is assumed that boron is 4-coordinted, as supported by the ATR-FTIR and NMR data, then a similar N.sub.C value can be calculated as that for Bioglass®. However, it should be noted that there are three main limitations to using this approach with sol-gel derived glasses, where: 1) The above calculation does not take into account the increased surface area and porosity; 2) not all boron is 4-coordinated; and 3) the sol-gel process results in residual OH.sup.− groups on the surface, which may contribute to the bioactivity rates of the SGBGs in this study, even in N.sub.C ranges where bioactivity is thought to be inhibited. The latter, has been previously demonstrated for sol-gel derived silicate-based glasses. A plot (FIG. 5) of the relationship between N.sub.C and the difference between T.sub.c and T.sub.g of each glass composition of Example 1, demonstrated a linear correlation (R.sup.2=0.9384; FIG. 5). An increase in the difference between T.sub.c and T.sub.g provides a good estimate of the tendency of the glass to remain amorphous, which can be used to predict the glass forming ability of the compositions of Example 1. As verified by XRD and ATR-FTIR (FIGS. 2 and 3, respectively), glasses with higher borate content remained amorphous at higher calcination temperatures and suggested that these compositions favored glass formation.

Example 8

Reactivity: Vapour Sorption of the Glasses

[0124] Under controlled temperature and humidity, vapour sorption of the biomaterials of Examples 1 and 2 was examined using a DVS Intrinsic (Surface Measurements Systems Ltd.) measuring mass changes (±0.1 μg). This may be an indicator of their potential solubility and reactivity. Approximately 5 mg of the glass particles from Examples 1 and 2 were placed in an aluminum pan and inserted into a chamber kept at 37±0.05° C. Two methods of analysis were carried out: 1) the relative humidity (RH) was increased stepwise at 5% RH up to 90% RH then back down to 0% RH while the relative mass change was measured when equilibrium was reached or after maximum of 4 h; 2) the glass particles were directly exposed to 90% RH for 6 h, which was then reduced back down to 0% RH for another 6 h.

[0125] As seen in FIG. 6, mass changes for all biomaterials of Example 1 were similar up to 60% RH and increased significantly until 90% RH. The % mass change at 90% RH correlated to the formulation with the lowest borate content, B36, having the greatest percent mass change (˜74%) compared to B61 (˜36%). Upon decreasing RH, desorption mainly occurred between 90 and 55% RH for the lower borate content glasses. The final mass change at 0% RH ranged between 19.5 and 23.7% and correlated with glass composition.

[0126] In contrast, the comparative glass of Example 2 did not indicate mass change until approximately 65% RH, where it gradually increased to ˜11% at 90% RH, while the biomaterials of Example 1 began to change at as little as 5% RH. Desorption in the comparative glass of Example 2 mainly occurred between 90 and 65% RH reaching a final mass change of ˜3%.

[0127] Upon the direct exposure to 90% RH for 6 hours, the biomaterial glasses of Example 1 experienced an immediate rapid increase in % mass change within the first 2 h, and followed by a slower rate of increase up to 6 h. The rate and extent of % mass change (˜58 to ˜41%) were dependent on the glass composition and increased with a decrease in glass borate content suggesting greater extent of bioactivity. Upon lowering the RH to 0%, the mass change % rapidly decreased with lower borate content glasses indicating greater extent of final % mass change. For the comparative glass of Example 2, its direct exposure to 90% RH resulted in ˜11% mass change at 6 h, which was less than that of the sol-gel equivalent (B46 of Example 2) (˜48%). In addition, beyond the first 2 h, 45B5 of Example 2 underwent a linear % mass change, similar to that experienced by melt-derived silicate and phosphate-based glasses.

[0128] Therefore, reactivity, as indicated by vapour sorption, was found to be highly dependent on SGBG composition, where atomic and molecular structures play prominent roles in the chemical durability of multi-component glasses and can be related to N.sub.C. It is likely that in the case of lower borate content glasses, the fewer boron units resulted in more terminal groups, specifically OH.sup.−, which were more prone to aqueous interaction and resulted in higher extents of mass change. This is particularly relevant to sol-gel derived glasses as these terminal groups are not fully removed during drying and calcination. On the other hand, the role of SGBG textural properties (SSA and pore volume) were prominent as B46 and 45B5 experienced drastically different % mass changes. While DVS is mainly used in the pharmaceutical and food science fields it has recently been used to measure the reactivity of bioactive phosphate glasses. This technique has shown to correlate well with weight loss measurements and since it is difficult to obtain accurate weight loss measurements with nano porous powders, the DVS serves as good substitute to examine aqueous reactivity.

Example 9

Reactivity: Ion Release

[0129] Ion release of B, Ca, Na, and P from the biomaterials of Example 1 compared to the glass of Example 2 in deionized water (DIW) at a 1.5 mg/mL ratio, were quantified using an inductively coupled plasma optical emission spectrophotometer (ICP-OES) (Thermo Scientific iCAP 6500). 10 mL aliquots were filtered through a 0.2 μm nylon filter and stored in a 15 mL falcon tube to which 4% (w/v) nitric acid (Fisher Scientific, CAN) was added. NIST standards were used as calibration values. The pH of the DIW solution was measured at each time point using an Accumet XL20 pH meter (Fisher Scientific).

[0130] ICP-OES measurements up to day 7 in DIW revealed the release of all four SGBG components (FIG. 7). There were rapid rates of release of boron and sodium ions, which were dependent on borate and soda contents in the Example 1 glasses, respectively. Beyond the 6 h time point, boron and sodium ion concentrations remained constant, indicating their full release. While the release rates of calcium and phosphorous ions were also rapid within the first 6 h, the extents of the release of these ions were inversely related to glass composition. Beyond the 6 h time point, there was a concomitant decrease in the concentrations of these ions in solution that was possibly due to their complexing. The rates of ion release from the Example 1 glass compositions were supported by changes in DIW pH, rapidly increasing within the first 6 h, then stabilising at a constant value, which suggested full ion release. The increase in pH values corresponded with glass composition, where glasses with lower borate content (i.e., higher soda content) resulted in greater extents of pH increase. For all Example 1 compositions, the starting pH was about 5.5 and had risen steeply by 6 h. For B36, the pH rose sharply to about 10, for B41 to about 9.6, for B46 to about 9.3, for B51 to about 9.1, for B56 to about 9.0, for B61 to about 8.8.

[0131] The ion release profiles from the Example 1 glasses were distinct from those of the comparative melt-derived glass of Example 2 (45B5), which demonstrated a slower, more gradual release rate, where by day 3, the concentration of boron ions in solution reached a similar level to that of B46 achieved after 6 h. Sodium ion release was also higher in 45B5, which may be due to the lower release of boron ions, leading to a more basic solution. The higher extent of sodium ion release resulted in higher pH values, even with the significantly lower textural properties of the melt-derived 45B5 (pH rising from about 5.5 to about 10.5 at 6 h. Furthermore, the release of calcium ions from 45B5 displayed a contrasting trend to that of B46, where after the 6 h time point, it steadily increased in concentration that eventually stabilized at day 3. The extent of phosphorous ion release from 45B5 was found to be at least 10-fold lower when compared to B46.

[0132] It is believed that the rate of calcium and phosphorus release from the biomaterial influences the rate and/or extent of mineralization.

Example 10

Bioactivity—In Vitro Mineralization

[0133] The extent of mineralization of the biomaterials of Example 1, calcined at 400° C., and the glass of Example 2, using Kokubo's simulated body fluid (SBF) (pH 7.4) was investigated. The use of SBF to examine bioactivity is regarded as the standard method to examine acellular mineralization. Glass powder was added to sterile 50 mL falcon tubes containing SBF at a 1.5 mg/mL ratio and stored in an oven at 37° C.±1° C. Twice per day, the vials were gently agitated in order to reduce agglomeration of the particles. Mineralization of the glasses was examined at the end of 6 h, 1 d, 3 d, and 7 d time points when the powders were gently rinsed twice with DIW then twice with ethanol, dried overnight at room temperature, and then dried in an oven for 1 d at 60° C. At each time point the pH of the SBF solution was measured using an Accumet XL20 pH meter (Fisher Scientific).

[0134] In a parallel study on the biomaterials of Example 1, to further examine in vitro mineralization, a 0.02M K.sub.2HPO.sub.4 solution, adjusted to pH 7 using dilute HCI was used for the same procedure as the SBF. This solution has commonly been used to investigate acellular mineralization of borate based glasses (A. Yao et al, J. American Ceramic Society 90, 303-306 (2007); W. Huang et al. J. Materials Science: Materials in Medicine 17, 583-596 (2006)).

[0135] ATR-FTIR confirmed carbonated-apatite was initiated after immersion in SBF for as little as 6 hours (FIG. 8). The strong band at ˜1018 cm.sup.−1 along with shoulders at 961 and 1062 cm.sup.−1, are characteristic of the bending modes v1 and v3 of PO.sub.4.sup.3− respectively. The broad bands at ˜1470 cm.sup.−1 and ˜1421 cm.sup.−1 are characteristic of the stretching mode (v1) and (v3) of CO.sub.3.sup.2− respectively. The weak band at 1640 cm.sup.−1 is due to the bending mode (v2) of water. The sharp peak at ˜870 cm.sup.−1 indicates the bending mode (v2) of CO.sub.3.sup.2− as traditionally seen in carbonated apatites. However this peak was also observed in the as made compositions of Example 1, particularly in lower borate content glasses, as the B—O stretching of boroxol rings suggesting a combination of both structural forms. An increase in exposure time to SBF corresponded with these peaks becoming sharper and more defined, indicating the progressive crystallization of HCA. While higher borate containing SGBGs presented more defined phosphate peaks at earlier time points, the spectra at day 7 indicated smaller carbonate peaks.

[0136] XRD analysis further confirmed apatite formation in all Example 1 glasses (FIG. 9) in 6 h as indicated by the appearance of peaks at ˜25 and ˜32° 2θ which are characteristic of hydroxyapatite formation (Joint Committee on Powder Diffraction Standards (JCPDS) 09-0432) where higher borate content glasses indicated more defined peaks. The peaks became more defined with time suggesting increased bone mineral formation. At day 1 all compositions showed more prominent broad hydroxycarbonate apatite (HCA) peaks, which may indicate nanosized or not fully crystallized HA. On the other hand, B36 showed the formation of a calcite phase (“□”, JCPDS 5-0586), attributable to its higher lime content, along with the lower N.sub.C likely resulting in calcite forming terminal groups. Furthermore, and in line with that observed in DIW, the increase in the pH of SBF solution corresponded with glass borate content, which favours HCA formation (FIG. 9). Without being limited to theory, the conversion mechanism of borate-based glasses to hydroxyapatite is thought to be similar to that of Bioglass® 45S5, but without the formation of an SiO.sub.2 rich layer. It is thought that in the case of borate-based glasses, an HCA is initially formed on the outer glass surface which then continually reacts towards its center, causing a reduction in volume, until full conversion has taken place. This has been attributed to the relatively rapid release of BO.sub.3.sup.3− and Na.sup.1 ions, while the remaining Ca.sup.2+ and PO.sub.4.sup.3− ions from the glass migrate to the surface and react with similar ions in the SBF solution leading to formation of an amorphous calcium-phosphate layer that eventually crystallizes into HCA. With the SGBG formulations investigated in this study, it is proposed that a similar mechanism of conversion occurs, yet at a significantly more rapid rate compared to melt-derived glasses, attributable possibly to their significantly higher textural properties and rapid ion release rates. Therefore, in order to further examine the effect of sol-gel processing on the bioactivity of borate-based glasses, the rates of mineralization of B46 and 45B5 were directly compared.

[0137] Increased pH of the mineralization media over time can also be a good indicator of HA formation. For all the formulations tested, the pH of the solution (SBF, DIW) increased (FIG. 10). The solid line in FIG. 10 represents the SBF data and the dashed line represents the deionized water data. The biomaterials of Example 1 rapidly reached their pH peak at 6 hours suggesting rapid ion exchange. Formulations with the highest Na content maintained the highest pH. The comparative melt-derived biomaterial of the same composition (Example 2) took about 7 days to reach the same pH level. This suggests that the dissolution rate of the biomaterials of the present disclosure is much more rapid than those with a lower surface area and pore volume.

[0138] The alkaline nature of the biomaterial solution precursors is thought to increase the connectivity of the glassy network and improve gelation. During processing this was observed as many of the compositions of Example 1 began to gel within 5 minutes of the final sodium addition.

[0139] The onset mineralization of all SGBG compositions occurred within 3 hours in SBF, demonstrating at least a ˜25 fold increase in bioactivity rate relative to melt-derived borate-based glasses. The ability of the SGBGs to rapidly convert to bone-like HCA holds promise for the repair and augmentation of mineralized tissues.

Example 11

Scanning Electron Microscopy

[0140] Scanning electron microscopy (SEM) was used to investigate the morphological properties of the glass powders. Samples were sputter coated with Au/Pd and analysis was performed with an Inspect F50 Field Emission Scanning Electron Microscope (FEI Corporation, USA) at 10 kV. To determine conversion of HCA, Energy Dispersive Spectroscopy (EDS) using an attached EDAX and a TEAM EDS Analysis System was performed at 20 kV on 8 unique glass surface areas to determine the Ca/P ratio at the 6 h and 7 d time points.

[0141] FIGS. 11a and 11b show SEM micrographs of B46 of Example 1 and 45B5 of comparative Example 2 at different time points: immediately after calcining, 6 hours in SBF and 7 days in SBF. The scale bar in FIG. 11 represents 10 μm and the inset scale bar represents 1 μm. Attributable to sol-gel processing, the surface of calcined B46 exhibited a rough, nanoporous texture; corroborating the textural properties in Table 4. In contrast, 45B5 displayed a relatively smooth surface appearance, typical of melt-derived glasses. However, surface roughness in both glasses became more apparent with time in SBF. EDAX analysis of B46 after 6 h in SBF indicated the rapid formation of an apatite-like calcium-phosphate layer, with a Ca/P ratio approaching that of HCA (1.60±0.01). This was in contrast to 45B5, where the Ca/P ratio at the 6 h time point was more reflective of the glass composition. However, by day 7 in SBF, a comparable Ca/P ratio was observed in both glasses. The rough porous surface of the calcined B46 becomes smoother after a 6 hour immersion in SBF possibly attributable to the washing of loose nanoparticles. However, the surfaces regained their textured appearance with time in SBF, and by day 7, the typically observed HCA morphology was apparent, which correlated with XRD and ATR-FTIR.

[0142] The pH change over time of formulation B46 of Example 1 and formulation 45B5 of Example 2 (comparative example) in three different dissolution media (deionised water (DIW); simulated body fluid (SBF); and potassium hydrogen phosphate (K2HPO4)) were investigated (FIG. 12). It was found that the melt-derived formulation of Example 2 has a higher pH than the sol-gel derived formulation of Example 1, regardless of the dissolution media.

[0143] ATR-FTIR spectroscopy of the glasses at earlier time points in SBF indicated that HCA-like formation was achieved in as little as 0.5 h in B46, compared to 3 days in the case of 45B5 (FIG. 13). Furthermore, XRD diffractograms showed that HCA formation was initiated after 3 h for B46, compared to 3 days for 45B5. This demonstration of the highly bioactive nature of the biomaterials of the present disclosure may be attributable to their ability to release greater extents of phosphate ions, thus favoring rapid HCA formation in SBF.

[0144] As a comparison to SBF, this study also investigated the mineralization of B46 and 45B5 in 0.02 M K.sub.2HPO.sub.4, which provided a 20 fold increase in phosphate content compared to SBF. It was found that apatite formation initiated within 6 h in both glasses, indicating that K.sub.2HPO.sub.4 artificially promotes rates of in vitro mineralization through the provision of excess, non-physiological concentrations of phosphate ions (FIG. 14). Furthermore, the appearance of a sharper PO.sub.4.sup.3− peak and more pronounced shoulder regions in the ATR-FTIR spectra, indicated that the exposure to K.sub.2HPO.sub.4 did not favor the production of CO.sub.3.sup.2− and OH.sub.3.sup.2− peaks, indicators of carbonated-apatite formation. Therefore, the use of SBF in assessing in vitro bioactivity can be regarded as more representative than K.sub.2HPO.sub.4 solution.

Example 12

Calcining Temperature

[0145] FIG. 15 demonstrates that the porosity and surface area of the biomaterials of the present disclosure can be controlled by firing temperature during the calcination process. The biomaterials become more dense with higher firing temperatures thus diminishing their nanopores resulting in small surface areas and total pore volume. This is supported by XRD and FTIR since at these higher temperatures the glasses become crystalline. Therefore, the surface texture properties and hence the reactivity of the biomaterials of the present disclosure can be controlled by processing temperature.

Example 13

Cell Viability

[0146] Mesenchymal stem cells (MSCs) were grown to confluence in a 96 well plate. The biomaterials of Example 1 were dissolved in Dulbecco's Modified Eagle Medium (DMEM) for one day at 37 C at a 50 mg/mL ratio. Aliquots of the media with dissolved ions were combined with new DMEM media at a 1:1 and 1:4 ratio then added to the cells at separate time points (2 h, 6 h, 10 h, and 24 h). The cells were then stained with Calcein-AM and fluorescence was spectroscopically read. The values were normalized to the control (wells with only media). The results (FIG. 16) showed that mesenchymal stem cells remained largely viable when in contact with the ionic release from the biomaterials of Example 1. The viability of the cells appeared to be related to composition and ionic concentration. From this it can be predicted that cellular behaviour in vivo can be controlled by controlling the dose of ionic release as well as the relative proportions of each ion.

Example 14

Borate-Glass Biomaterial without Phosphates

[0147] A three component borate-based glass biomaterial was made based on boron-sodium-calcium oxides, with borate as the sole network forming component. Specifically, the biomaterial made had the following composition: 54 wt % B.sub.2O.sub.3—22 wt % CaO—24 wt % Na.sub.2O. The method here differed from that of Example 1 in that no phosphate precursor was used, and 6.53 g boric acid was dissolved in 58.33 mL 100% EtOH, followed by mixing with 25 g calcium methoxyethoxide (20%) and 11.36 g of sodium methoxide (25%). The solution was observed to have gelled within 30 minutes of adding the final precursor.

[0148] It should be appreciated that the invention is not limited to the particular embodiments described and illustrated herein but includes all modifications and variations falling within the scope of the invention as defined in the appended claims.