The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
This office action is in response to Applicants’ amendments/remarks received on July 20, 2026.
Rejections and/or objections not reiterated from previous office actions are hereby withdrawn.
Claims 17-18 are canceled. Claims 1-16 are under consideration.
Priority: This application is a 371 of PCT/US2022/048912, filed November 4, 2022, which claims benefit of provisional application 63/275565, filed November 4, 2021.
Objections and Rejections
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
Claims 1-2, 8-11, 13 are rejected under 35 U.S.C. 103 as being unpatentable over Schaffner et al. (2017 Sci Adv 3(12):eaao6804, 9 pages; previously cited) in view of Qian et al. (US 20200109299; IDS 05.03.24, previously cited) and Hinton et al. (2015 Sci Adv 1:e1500758, 10 pages). Schaffner et al. disclose 3D printing of bacteria into functional complex materials by embedding bacteria in a biocompatible and functionalized 3D printing ink and printing “living materials” or “Flinks” (at least p. 1-2, also Fig. 1). Schaffner et al. disclose a method for 3D printing of Flinks comprising providing an ink composition comprising sodium hyaluronate (HA) functionalized with glycidyl methacrylate (GM), a solvent, bacteria, and Irgacure 2959 (a photoinitiator) (at least p. 7-8), 3D printing a pattern in a hydrogel support matrix using the ink composition in a container, and forming a Flink material comprising the living bacteria by curing the printed material (at least p. 2 Fig. 1, p. 5-6 Fig. 3-4, p. 7-8). Schaffner et al. disclose printing various patterns and/or shapes of hydrogels containing bacteria, including printing a plurality of separate hydrogel shapes in one container and a multimaterial hydrogel (at least p. 5, Fig. 3). Schaffner et al. disclose complex materials can be assembled by the 3D printing ink comprising bacteria (p. 1), allowing the possibility to combine different organisms and chemistries in a single process, allowing for digital shaping of living materials into new geometries and adaptive functional architectures (p. 2), including for printing biocompatible 3D cellular structures for bioremediation and complex shaped tissue for biomedical applications (at least p. 1-2). Schaffner et al. differ from the method of instant claim 1 by not explicitly teaching a monomer and a hydrogel support matrix.
Qian et al. also disclose methods for making a living structure from a bio-ink material of living freeze-dried cells, the bio-ink material comprising fillers, binders, and a photoinitiator (at least abstract, paragraphs 0148-0170). Qian et al. disclose in some embodiments, the bio-ink comprises the cells and more than one filler component (at least paragraphs 0062-0063), the filler further comprises a binder, the binder is an oligomer, monomer, or mixtures thereof, where the filler further comprises a photo-initiator that absorbs light and initiates photopolymerization of the binder material (i.e. the oligomer and/or monomer) (at least paragraph 0064).
Hinton et al. disclose the three-dimensional printing of complex biological structures by freeform reversible embedding of suspended hydrogels (p. 1). Hinton et al. disclose the structures are built by embedding the printed hydrogel within a secondary hydrogel that serves as a temporary biocompatible support (at least p. 1-2). Hinton et al. disclose the development of a 3D bioprinting technique termed freeform reversible embedding of suspended hydrogels (FRESH) (at least p. 1-2). Hinton et al. disclose that the key innovation of FRESH is deposition and embedding of the hydrogel(s) being printed within a second hydrogel support bath that maintains the intended structure during the print process and significantly improves print fidelity (at least p. 2, also Fig. 1).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to incorporate a polymerizable monomer of Qian et al. for the polymer in a plurality of ink compositions provided in the method of 3D printing a living material comprising bacteria of Schaffner et al. noted above and to further 3D print a plurality of patterns or structures with the ink compositions of Schaffner et al. in view of Qian et al. into a hydrogel support as disclosed in Hinton et al., to thereby arrive at the claimed method for 3D printing living materials comprising providing a plurality of ink compositions comprising a polymerizable monomer, a crosslinking agent, a photoinitiator, and a solvent, 3D printing a plurality of patterns or structures in a hydrogel support matrix using the ink compositions in a container, and forming a 3D printed living material comprising the living bacteria by curing the printed living material (instant claim 1). The motivation to do so is given by the prior art, which disclose ink compositions for 3D printing comprises materials including monomers, polymers, and/or oligomers and where the 3D printing is in a hydrogel support. One of ordinary skill would have a reasonable expectation of success because the prior art discloses materials for ink compositions utilized in 3D printing of living materials are known.
Regarding instant claim 2, Schaffner et al. disclose bacteria is in the printed 3D living material (at least p. 1-2, also Fig. 1) and Qian et al. also disclose cells in the living material are microbes, in some embodiments, the microbes include bacteria and are E. coli, or the microbes include yeast and are S. cerevisiae (at least paragraph 0053). Therefore, it would be obvious to one of ordinary skill that the bacteria in the ink composition for 3D printing a living material is selected to be E. coli.
Regarding instant claim 8, Schaffner et al. disclose the photoinitiator Irgacure 2959 is 2-hydroxy-4’-(2-hydroxyethoxy)-2-methylpropiophenone (p. 7) and Qian et al. also disclose suitable photo-initiators include Irgacures (at least paragraph 0064). Therefore, it would be obvious to one of ordinary skill to arrive at the recited 2-hydroxy-4’-(2-hydroxyethoxy)-2-methylpropiophenone photoinitiator in the ink composition for 3D printing a living material.
Regarding instant claim 9, Schaffner et al. disclose mixing the components to form the ink composition (at least p. 7) and Qian et al. also disclose mixing the components to form the bio-ink composition (at least paragraphs 0111-0115). Therefore, it would be obvious to one of ordinary skill to mix the components of the ink composition comprising a polymerizable monomer, cross-linking agent, photoinitiator, and solvent of Schaffner et al. in view of Qian et al. and Hinton et al. noted above.
Regarding instant claims 10-11, Qian et al. disclose in some embodiments, the bio-ink comprises more than one filler component, where the filler component is alginate (at least paragraph 0061). Therefore, it would be obvious to one of ordinary skill to further include an additional polymer, the polymer being alginate, in the ink composition comprising a polymerizable monomer, cross-linking agent, photoinitiator, and solvent noted above.
Regarding instant claim 13, Schaffner et al. disclose curing the Flinks comprising photoinitiator with light (at least p. 7) and Qian et al. disclose activating the photoinitiator by irradiation with ultraviolet (UV) light (at least paragraph 0064). Therefore, it would be obvious to one of ordinary skill to arrive at the recited curing the 3D printed pattern with at least one wavelength of light to activate the photoinitiator.
Reply: Applicants’ amendments/remarks have been considered but they are not persuasive. The claims remain unpatentable under 103 over Schaffner et al. in view of Qian et al. and newly cited Hinton et al. for the reasons noted above and herein.
Applicants assert that first, none of the cited art teaches or suggests printing multiple inks/layers of a 3D structure and then having a single curing step. Applicants assert that the light-curable Flinks were cross-linked in a layer-by-layer fashion using an OmniCure S1000; each layer was illuminated for 60s (Schaffner et al. p. 7). Applicants assert that further, when Schaffner et al. disclose the printing of different materials, they were still printed on different layers. Applicants assert that these are different layers that would be cured sequentially as noted in Schaffner et al.
Applicants’ remarks are not persuasive. Schaffner et al. disclose that the 3D printing of ink compositions comprising microorganisms are demonstrated for both bioremediation and biomedical applications (at least p. 2). Schaffner et al. disclose 3D printing various patterns and/or shapes of hydrogels containing bacteria, including printing a plurality of separate hydrogel shapes in one container and a multimaterial hydrogel (at least p. 5, Fig. 3). Schaffner et al. further disclose that using multimaterial DIW (direct ink writing) of Flinks, bacteria can be incorporated and grown in specific regions of printed structures with high accuracy and freedom of shape (Fig. 3, A to C). Schaffner et al. disclose that full control of bacteria local concentration is possible by multimaterial DIW because each of the cartridges can be loaded with different bacterial strains at various concentrations and eventually extruded at any point in the object (at least p. 3). Therefore, Schaffner et al. reasonably disclose providing a plurality of ink compositions for 3D printing a plurality of 3D hydrogel structures comprising embedded microorganisms and curing the 3D hydrogel structures.
While Applicants assert that Schaffner et al. disclose the light-curable Flinks were cross-linked in a layer-by-layer fashion using an OmniCure S1000; each layer was illuminated for 60s (Schaffner et al. p. 7), it is noted that Schaffner et al. disclose this layer-by-layer fashion for 3D printing Flinks for bioremediation in a 3D-printed lattice (p. 2, 7). Schaffner et al. do not disclose that the light-curable Flinks when 3D printed into a plurality of other types of complex shapes and/or patterns for other types of applications, i.e. biomedical applications (p. 3, 5, also Fig. 3C), necessarily need to be cured in a layer-by-layer fashion. It would be reasonable and obvious to one of ordinary skill that the plurality of 3D printed hydrogel shapes printed in one container disclosed for instance in Fig. 3C of Schaffner et al. can be illuminated and cured together in a single curing step in the container.
Therefore, Applicants’ remarks that Schaffner et al. do not disclose printing a plurality of ink compositions are not persuasive.
Applicants assert that second, the claim requires printing the pattern into a hydrogel support matrix held in a container. Applicants submit that neither Schaffner et al. nor Qian et al. disclose this.
Applicants’ remarks are not persuasive. It is noted that instant claim 1 does not actually recite printing the pattern into a hydrogel support matrix. The instant claim recites printing the pattern in a hydrogel support matrix, which can reasonably be interpreted as the ink composition being printed is a hydrogel support. Nevertheless, Schaffner et al. is now cited with Hinton et al., which disclose 3D printing a plurality of patterns into a hydrogel support to embed hydrogel(s) with a second hydrogel support (at least p. 2, also Fig. 1). Schaffner et al. disclose that the deposition and embedding of the hydrogel(s) being printed within a second hydrogel support bath maintains the intended structure during the print process and significantly improves print fidelity (at least p. 2, also Fig. 1).
Therefore, it would have been obvious to incorporate a polymerizable monomer of Qian et al. for the polymer in a plurality of ink compositions provided in the method of 3D printing a living material comprising bacteria of Schaffner et al. noted above and to further 3D print a plurality of patterns or structures with the ink compositions of Schaffner et al. in view of Qian et al. into a hydrogel support as disclosed in Hinton et al., to thereby arrive at the claimed method for 3D printing living materials comprising providing a plurality of ink compositions comprising a polymerizable monomer, a crosslinking agent, a photoinitiator, and a solvent, 3D printing a plurality of patterns or structures in a hydrogel support matrix using the ink compositions in a container, and forming a 3D printed living material comprising the living bacteria by curing the printed living material (instant claim 1).
For at least these reasons, the 103 rejection is maintained.
Claims 1-2, 6-7, 8-11, 13 are rejected under 35 U.S.C. 103 as being unpatentable over Schaffner et al. (2017 Sci Adv 3(12):eaao6804, 9 pages; previously cited) in view of Qian et al. (US 20200109299; IDS 05.03.24, previously cited), Hinton et al. (2015 Sci Adv 1:e1500758, 10 pages), and Zhu et al. (2020 Sci Adv 6:eaba5575, 10 pages; previously cited). The teachings of Schaffner et al., Qian et al., and Hinton et al. over at least instant claims 1-2, 8-11, 13 are noted above.
Regarding instant claims 6-7, Qian et al. disclose that the bio-ink composition comprises a polymerizable monomer (at least paragraph 0064). Zhu et al. disclose an ink composition comprising acrylamide as the monomer, N,N’-methylenebisacrylamide as the cross-linker, and a photo-initiator (at least p. 8). Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to incorporate the acrylamide and N,N’-methylenebisacrylamide of Zhu et al. for the monomer and crosslinking agent in the ink composition of Schaffner et al. in view of Qian et al. and Hinton et al. noted above in the method of 3D printing a living material, to thereby arrive at the claimed method for 3D printing living materials comprising providing a plurality of ink compositions comprising a acrylamide, N,N’-methylenebisacrylamide, a photoinitiator, and a solvent, 3D printing a plurality of patterns or structures in a hydrogel support matrix using the ink compositions in a container, and forming a 3D printed living material comprising the living bacteria by curing the printed living material (instant claims 6-7). The motivation to do so is given by the prior art, which disclose ink compositions for 3D printing comprises materials including monomers, where acrylamide is a known monomer for preparing 3D ink. One of ordinary skill would have a reasonable expectation of success because the prior art discloses materials for ink compositions utilized in 3D printing are known.
Reply: Applicants’ amendments/remarks have been considered but they are not persuasive. The reasons for maintaining Schaffner et al. are the same as noted above.
Claims 1-2, 3-5, 8-11, 13 are rejected under 35 U.S.C. 103 as being unpatentable over Schaffner et al. (2017 Sci Adv 3(12):eaao6804, 9 pages; previously cited) in view of Qian et al. (US 20200109299; IDS 05.03.24, previously cited), Hinton et al. (2015 Sci Adv 1:e1500758, 10 pages), and Millik et al. (2019 Biofabrication 11:045009, 11 pages; previously cited). The teachings of Schaffner et al., Qian et al., and Hinton et al. over at least instant claims 1-2, 8-11, 13 are noted above.
Regarding instant claims 3-4, Qian et al. disclose the bio-inks comprise a cell density of at least 50 wt% (at least paragraph 0073), in some embodiments, a sample of cells are obtained from a source, isolated, and dried, where prior to drying, the sample of cells is concentrated, by for instance, centrifuging (at least paragraph 0074). Qian et al. disclose the bio-ink comprising the cell granules, solvent, polymer, and photoinitiator, can be loaded into a syringe barrel for printing (at least paragraph 0113). Millik et al. disclose 3D printing for hydrogels (at least p. 1). Millik et al. disclose that prior to using the hydrogel in 3D printing, remaining bubbles are eliminated by centrifugation (at least p. 3-4). Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention that the ink composition of Schaffner et al. in view of Qian et al. and Hinton et al. noted above can be formed by mixing a pellet having a known cell amount with the ink composition components, centrifuging the ink composition, and loading the centrifuged ink composition into a syringe for injection (instant claims 3-4). The motivation to do so is given by the prior art, which disclose that centrifuging concentrates the components of a composition and eliminates air bubbles. One of ordinary skill would have a reasonable expectation of success because methods and conditions for preparing bioink compositions for 3D printing living materials are known in the prior art.
Regarding instant claim 5, as noted above, Qian et al. disclose that the sample of cells are obtained from a source and centrifuged (at least paragraph 0074). Schaffner et al. disclose that the cells for incorporating or embedding in the bio-ink composition are grown in culture medium (at least p. 8). Schaffner et al. also disclose cultures can be pelleted by centrifugation and suspended in MM for ink preparation or direct inoculation (at least p. 8). Therefore, it would have been obvious to one of ordinary skill that the cells for incorporation into the bioink composition can be prepared in pellet form by inoculating cells into a culture or growth medium, growing the cells in the culture medium, and centrifuging the cells to form a cell pellet (instant claim 5). One of ordinary skill would have a reasonable expectation of success because methods and conditions for preparing bioink compositions for 3D printing living materials are known in the prior art.
Reply: Applicants’ amendments/remarks have been considered but they are not persuasive. The reasons for maintaining Schaffner et al. are the same as noted above.
Claims 1-2, 8-11, 12, 13 are rejected under 35 U.S.C. 103 as being unpatentable over Schaffner et al. (2017 Sci Adv 3(12):eaao6804, 9 pages; previously cited) in view of Qian et al. (US 20200109299; IDS 05.03.24, previously cited), Hinton et al. (2015 Sci Adv 1:e1500758, 10 pages), and Shoseyov et al. (US 20200179562; IDS 05.03.24, previously cited). The teachings of Schaffner et al., Qian et al., and Hinton et al. et al. over at least instant claims 1-2, 8-11, 13 are noted above.
Regarding instant claim 12, Qian et al. disclose that the bio-ink further comprises a hydrogel, where non-limiting examples of hydrogels include alginate (at least paragraph 0070). Qian et al. also disclose that the living structure is biocompatible with a physiological environment without eliciting unwanted and/or adverse effects to the environment (at least paragraph 0079). Shoseyov et al. also disclose material formulations for additive manufacturing or 3D printing (abstract), where 3D printing uses biological materials, optionally in combination with chemicals and/or cells (at least paragraphs 0013), where to allow for control on the curing, the building material commonly includes polymerizable moieties or groups that polymerize upon being dispensed, to preserve the geometric shape and provide the necessary physical properties of the final product (at least paragraph 0016). Shoseyov et al. disclose the receiving medium for the 3D printing is a supporting medium and can be biocompatible material and comprises a hydrogel (at least paragraphs 0117-0120). Shoseyov et al. also disclose the hydrogel is biocompatible and is such that when a biological moiety is impregnated or accumulated therein, an activity of the biological moiety is maintained (at least paragraph 0326), and where the hydrogel properties are governed by factors including the aqueous media content and composition (at least paragraph 0316). Shoseyov et al. disclose aqueous carriers (or media) comprising a culturing medium (at least paragraph 0296). Hinton et al. disclose that in preparing the hydrogel support receiving the 3D printing, the medium slurry is vortexed and centrifuged and repeated until no bubbles are observed (p. 7). Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to further incorporate forming a hydrogel composition by mixing growth medium that is sterile with a thickening agent (i.e. alginate); eliminating bubbles by centrifuging the hydrogel composition; and loading the hydrogel composition into a container for printing in the method of 3D printing a living material of Schaffner et al. in view of Qian et al. and Hinton et al. noted above (instant claim 12). The motivation to do so is given by the prior art, which disclose bioink compositions can be 3D printed into a hydrogel composition that is biocompatible for the embedded cells; therefore, it would be obvious that a biocompatible hydrogel composition reasonably comprises aqueous medium, including culture medium that is sterile, since the hydrogel functions to maintain a biocompatible environment while maintaining cell activity. One of ordinary skill would have a reasonable expectation of success because methods and conditions for preparing bioink compositions for 3D printing living materials are known in the prior art.
Reply: Applicants’ amendments/remarks have been considered but they are not persuasive. The reasons for maintaining Schaffner et al. are the same as noted above.
Claims 1-2, 8-11, 13, 14-16 are rejected under 35 U.S.C. 103 as being unpatentable over Schaffner et al. (2017 Sci Adv 3(12):eaao6804, 9 pages; previously cited) in view of Qian et al. (US 20200109299; IDS 05.03.24, previously cited), Hinton et al. (2015 Sci Adv 1:e1500758, 10 pages), Shoseyov et al. (US 20200179562; IDS 05.03.24, previously cited), and Wang et al. (2018 J Biomed Mater Res Part A:106A: 865-875; previously cited). The teachings of Schaffner et al., Qian et al., and Hinton et al. over at least instant claims 1-2, 8-11, 13, and the teachings of Shoseyov et al. are noted above.
Regarding instant claim 14, Shoseyov et al. disclose that for all technologies, the most important parameter determining the accuracy and efficiency of printing is the static and physical properties of the dispensed building materials, including viscosity and shear-thinning properties (at least paragraph 0018). Shoseyov et al. disclose in some embodiments, material formulations features shear-thinning behavior and/or thermal-thinning behavior (at least paragraphs 0300-0304), where shear-thinning property describes a behavior of a fluidic material that is reflected by a decrease in its viscosity, where thermal-thinning property describes a property of a fluidic material that is reflected by a decrease in its viscosity (at least paragraphs 0302-0303). Hinton et al. also disclose releasing the embedded hydrogel structure 3D printed in the hydrogel support by heating to 37º C and melting the gelatin (i.e. reducing the viscosity of the receiving hydrogel support (at least p. 2, Fig. 1). MPEP 2144.04 notes that design changes are obvious. In this instance, the prior art discloses that the viscosity of hydrogels printed with bioinks can be altered and optimized. Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to further reduce a viscosity of the receiving hydrogel support embedded with the 3D printed hydrogel comprising living bacteria in the method of 3D printing a living material of Schaffner et al. in view of Qian et al. and Hinton et al. noted above, to thereby release the living material (instant claim 14), as a matter of design choice. The motivation to do so is given by the prior art Shoseyov et al., which disclose that reduced viscosity is a recognized property for 3D printed hydrogels using bioink and Hinton et al., which also disclose melting the hydrogel support releases the embedded hydrogel structure 3D printed in the hydrogel support by heating. One of ordinary skill would have a reasonable expectation of success because the prior art discloses materials for ink compositions utilized in 3D printing of living materials are known.
Regarding instant claims 15-16, as noted above, Shoseyov et al. disclose that reduced viscosity is a recognized property for 3D printed hydrogels using bioink. Hinton et al. also disclose releasing the embedded hydrogel structure 3D printed in the hydrogel support by heating to 37º C and melting the gelatin (i.e. reducing the viscosity of the receiving hydrogel support (at least p. 2, Fig. 1). Wang et al. disclose that hydrogels fabricated by 3D printing can be eroded by PBS and at 37º C (at least p. 867). Therefore, it would have been obvious to one ordinary skill in the art to reduce the viscosity of the hydrogel support embedded with the 3D printed hydrogel comprising living bacteria in the method of 3D printing a living material of Schaffner et al. in view of Qian et al. and Hinton et al. noted above, comprising contacting at least a part of the hydrogel support with a buffered solution (PBS) (instant claim 15). The motivation to do so is given by the prior art. Shoseyov et al. disclose that reduced viscosity is a recognized property for 3D printed hydrogels and Wang et al. disclose hydrogel viscosity can be reduced by contact with a buffered solution. One of ordinary skill would have a reasonable expectation of success because the prior art discloses materials for ink compositions utilized in 3D printing are known. Further regarding instant claim 16, “[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation.” In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955). MPEP 2144.05. As noted above, Wang et al. disclose that hydrogels fabricated by 3D printing can be eroded by PBS (at least p. 867). Therefore, it would have been obvious to arrive at the recited 10X phosphate buffered solution by routine optimization. One of ordinary skill would have a reasonable expectation of success because the prior art discloses materials for ink compositions utilized in 3D printing are known.
Reply: Applicants’ amendments/remarks have been considered but they are not persuasive. The reasons for maintaining Schaffner et al. are the same as noted above.
No claim is allowed.
THIS ACTION IS MADE FINAL. Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/Marsha Tsay/Primary Examiner, Art Unit 1656
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