Methods of manufacturing injectable microgel scaffolds
Disclosed herein are methods of manufacturing injectable microgel scaffolds, including methods of producing, purifying and concentrating microgel particles therein. The microgel scaffolds of the present disclosure are useful for a wide range of applications, such as stabilizing an implanted medical device in an implant site in a subject. The microgel scaffolds are fluidic during application and annealed or crosslinked after application to the implant site in the subject. The microgel scaffolds may contain various therapeutic agents, including antibiotics and analgesics, throughout the gel.
1. A method comprising:
a) providing a membrane filtration system comprising a solid support with pores, each pore having a diameter of at most 9 micrometers (μm);
b) transferring dispersed microgel particles from a first continuous phase to a final continuous phase by controlled addition of an intermediate continuous phase to the first continuous phase, wherein the final continuous phase is immiscible with the first continuous phase;
c) maintaining a single miscible continuous phase containing the dispersed microgel particles;
d) applying the single miscible continuous phase containing the dispersed microgel particles to a membrane of the membrane filtration system; and
e) removing an impurity from the dispersed microgel particles or the single miscible continuous phase using size exclusion filtration by the membrane filtration system, thereby producing purified microgel particles.
2. The method of claim 1 , wherein transferring of step (b), the maintaining of step (c), the applying of step (d), and the removing of step (e) occur substantially simultaneously.
3. The method of claim 1 , wherein maintaining the single miscible phase is required for the membrane filtration system to remove the impurity from the dispersed microgel particles.
4. The method of claim 1 , wherein applying the single miscible phase to the membrane comprises applying the single miscible phase in a direction that is tangential to the membrane.
5. The method of claim 1 , wherein the membrane filtration system is selected from tangential flow filtration (TFF), ultrafiltration-diafiltration (UFDF), microfiltration-diafiltration (MFDF), or hollow-fiber-diafiltration (HFDF).
6. The method of claim 1 , wherein the first continuous phase comprises a solvent that is a non-polar oil.
7. The method of claim 1 , wherein the intermediate continuous phase comprises a solvent that is an alcohol solution.
8. The method of claim 1 , wherein the impurity is a surfactant.
9. The method of 1 , wherein the purified microgel particles comprise a backbone polymer and annealing components.
10. The method of claim 9 , further comprising producing a stabilized scaffold by introducing an annealing agent to the purified microgel particles, the annealing agent linking the annealing components of the purified microgel particles together to form the stabilized scaffold, the stabilized scaffold comprising pores having a median diameter of about 10 μm to about 35 μm.
11. The method of claim 10 , wherein between or about 10-40% of a total volume of the stabilized scaffold is made up of the pores.
12. The method of claim 9 , wherein annealing components each comprise, independently, a functional group selected from the group consisting of a vinyl sulfone, thiol, amine, imidazole, aldehyde, ketone, hydroxyl, azide, alkyne, vinyl, alkene, maleimide, carboxyl, N-hydroxysuccinimide (NETS) ester, isocyanate, isothiocyanate, hydroxylamine, and thione.
13. The method of claim 9 , wherein the annealing components each comprise, independently a reactive moiety selected from the group consisting of a catechol, a sialic acid, a boronic acid, a molecular cage, adamantane, biotin, and streptavidin.
14. The method of claim 10 , wherein linking comprises a reaction selected from the group consisting of Michael addition, amide bond coupling, Diels-Alder cycloaddition, Huisgen 1,3-dipolar cycloaddition, reductive amination, carbamate linkage, ester linkage, thioether linkage, disulfide bonding, hydrazone bonding, oxime coupling, and thiourea coupling.
15. The method of claim 10 , wherein linking the annealing components is performed by forming a covalent bond between at least two of the annealing components.
16. The method of claim 10 , wherein the annealing components comprise a first annealing component and a second annealing component, and wherein the first annealing component and the second annealing component are not the same.
17. The method of claim 16 , wherein there is at least 1% more of the second annealing component than the first annealing component in the stabilized scaffold.
18. The method of claim 16 , wherein a concentration of the first annealing component and a concentration of the second annealing component in the stabilized scaffold are not the same.
19. The method of claim 10 , wherein producing a stabilized scaffold is performed in vivo, at a site of an implanted medical device.
20. The method of claim 1 , wherein the final continuous phase comprises a solvent that is water.