DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 09-11-2025 has been entered.
Applicant's amendments to the claims and arguments filed on 09-11-2025 have been received and entered. Claims 1-5, 8 are pending in the instant application. Claims 1-5, 8 are under consideration.
Priority
This application is a 371 of PCT/KR2021/018256 filed on 12/03/2021. It is noted that there is no claim for foreign priority. Thus, the effective filing date of the instant application is the filing date of 12-03-2021.
Withdrawn - Claim Rejections - 35 USC § 112
Claim 8 was rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention. In view of Applicants' amendment of claim 8, the previous rejections of claims are hereby withdrawn. Applicants' arguments with respect to the withdrawn rejections are thereby rendered moot.
Withdrawn-Claim Rejections - 35 USC § 103
Claims 1, 2, 3 were rejected under 35 U.S.C. 103 as being unpatentable over Kang et al (KR 20180125776 A, Date Published: 2018-11-26) in view of Zhou et al (International Journal of Bioprinting, vol.3(2): 130–137. Doi: 10.18063/IJB.2017.02.003, July 11, 2017) and McCormack et al (Trends in Biotechnology, June 2020, Vol. 38, No. 6; Doi: 10.1016/j.tibtech.2019.12.020). In view of Applicants' amendment of base claim 1, the previous rejections of claims are hereby withdrawn. Applicants' arguments with respect to the withdrawn rejections are thereby rendered moot. The claims are however subject to new rejections over the prior art of record, as set forth below.
Claim 4 was rejected under 35 U.S.C. 103 as being unpatentable over Kang et al (KR 20180125776 A, Date Published: 2018-11-26) in view of Zhou et al (International Journal of Bioprinting, vol.3(2): 130–137. Doi: 10.18063/IJB.2017.02.003, July 11, 2017) and McCormack et al (Trends in Biotechnology, June 2020, Vol. 38, No. 6; Doi: 10.1016/j.tibtech.2019.12.020) as applied to claims 1, 2, 3 above, and further in view of Zhuang et al (PLoS ONE 14(6): e0216776. Doi: 10.1371/journal.pone.0216776, June 12, 2019). The rejection is withdrawn for the reasons discussed above.
Claims 5, 8 were rejected under 35 U.S.C. 103 as being unpatentable over Kang et al (KR 20180125776 A, Date Published: 2018-11-26) in view of Zhou et al (International Journal of Bioprinting, vol.3(2): 130–137. Doi: 10.18063/IJB.2017.02.003) and McCormack et al (Trends in Biotechnology, June 2020, Vol. 38, No. 6; Doi: 10.1016/j.tibtech.2019.12.020) as applied to claims 1, 2, 3 above, and further in view of Ahn et al (Pub. No.: US 2019/0142995 A1, Pub . Date: May 16 , 2019). The rejection is withdrawn for the reasons discussed above.
New- Claim Rejections - 35 USC § 103 - necessitated by amendments
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, 3 are rejected under 35 U.S.C. 103 as being unpatentable over Kang et al (KR 20180125776 A, Date Published: 2018-11-26) in view of Skardal et al (Pub. No.: US 2020/0108172 A1, Pub. Date : Apr. 9, 2020) and McCormack et al (Trends in Biotechnology, June 2020, Vol. 38, No. 6; Doi: 10.1016/j.tibtech.2019.12.020).
Claim interpretation:
The specification of the claimed invention teaches that the alignment of cells means that cells are stretched above a predetermined level and have directionality ([0003], page 1). Forming output product in which cells are three dimensionally aligned: …… myoblasts of a mouse, were added thereto and suspended to produce a bio-ink. The above-produced bioink was mounted on an output barrel of a 3D bioprinter, and the bio-ink was outputted at a pressure of 60 kPa while moving a 25-gauge (G) nozzle in the path as illustrated in FIG. 1 to form an output product having a pattern as shown in FIG. 1 on a culture dish ([0043]-[0044], page 11). Thus, using bioink on a 3D bioprinter to form an output product having a pattern is interpreted as meeting the limitation of “aligning and culturing cells in three dimensions”.
The specification of the claimed invention teaches that the curing refers to a process in which a hydrogel in the output product is cross-linked by a covalent or noncovalent bond, and if necessary, according to the type of the hydrogel, a material which induces or promotes the crosslinking as described above may be used as a curing agent ([0031], page 7-8). Thus, curing the output product is interpreted as cross-linking by a covalent or noncovalent bond curing agent.
The specification of the claimed invention teaches that the hydrogel may be obtained by dispersing one dispersoid or a mixture of two or more dispersoids, such as gelatin, gellan gum, collagen, agar, alginic acid, hyaluronic acid, carrageenan, gum arabic, gum ghatti, pullulan gum, mannan gum, locust bean gum, and xanthan gum, in a dispersion medium containing water ([0024], Page 5, lines 18-23). Thus, the above polymers are interpreted as dispersoids.
Regarding to claim 1, the preamble, Kang et al teaches bioink composition for 3D printing. The bioink composition comprising a decellularized extracellular matrix (dECM) powder and hydrogel. Also, Kang et al stated that the present invention can be very useful to produce an artificial tissue or a tumor model using 3D bio-printing, and it can be usefully applied to tissue engineering scaffolds, cell-based sensors, and drug/toxicity screening. (Abstract)
Regarding to claim 1, the claimed: mixing cells with a hydrogel comprising unmodified gelatin as a dispersoid to prepare a bio-ink, Kang et al teaches that in order to produce extracellular matrix (dECM) -based bio-ink with improved printability, gelatin-based bio-ink (hydrogel) was first prepared (Example 1-1, page 4). To evaluate the cytocompatibility of the extracellular matrix-based bio-ink prepared in Example 1-4 with respect to the cells, dECM-based bio-ink was added at a concentration of 0.5x 106 cells / NIH3T3 cells (fibroblasts) were encapsulated (Example 2, page 6 )
Regarding to claim 1, the claimed: outputting the bio-ink while moving a nozzle of a discharge device to form an output product having a pattern having at least parallel portions, Kang et al teaches that to evaluate the printability and the cell viability of the dECM based bio-ink prepared in Example 1-4, 3-D bio-printers (3-D bioprinter, home-grown) was measured for cell survival (viability). Specifically, a printing test of a dECM-based bio ink was performed in an 8 x 4 mm pattern (having at least parallel portions, see Figure 9A and 9E below) using a 250 µm nozzle under a temperature condition of 18 oC. At this time, a gelatin-based bio-ink without a depleted outer cell matrix powder was used as a control group. As shown in Fig. 9, dECM-based bio-ink was stacked in four layers 9 A to C) was excellent in printability. In addition, viability of NIH3T3 cells in dECM-based bioinf after one day of printing was analyzed, and it was confirmed that cell viability after printing was superior to gelatin-based bio ink (Example 3, page 6-7, and Figure 9, also see Figure 6)
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Regarding to claim 1, the claimed: curing the output product by treating the output product with a solution containing transglutaminase or glutaraldehyde to make the gelatin crosslinked, Kang et al teaches that the in order to perform cross-linking according to the temperature, the cell-based bio-ink (dECM based bio-ink) and the cartridge into which the cells were injected were placed on ice for about 10 minutes and cross-linking. After printing, the three-dimensional structure was treated with 10U/ml of thrombin solution and then subjected to secondary crosslinking at room temperature (RT) for about 30 minutes (Example 4, page 7).
However, Kang et al do not teach a solution containing transglutaminase or glutaraldehyde to make the gelatin crosslinked. Skardal et al cure the deficiency.
Skardal et al teach gelatin nanoparticles including their use in a composition that may be in the form of a hydrogel (Abstract), and a composition of the present invention may be referred to as a “bioink” or a “ bioink composition” (both of which are used interchangeably herein ), and may comprise one or more live cell(s) ([0040], page 4). Skardal et al teach “One or more (e.g., 1, 2 , 3 , 4 , 5, 6 , 7, or more) additional components may be present in a composition of the present invention. For example, in some embodiments, a composition of the present invention comprises …… unmodified gelatin….” ([0061], page 6). Skardal et al teach the gelatin nanoparticles are crosslinked, wherein the gelatin nanoparticles are crosslinked with glutaraldehyde ([0007], page 1). In some embodiments, a gelatin nanoparticle of the present invention is crosslinked . In some embodiments , gelatin nanoparticle of the present invention is crosslinked with a crosslinking agent such as, but not limited to, glutaraldehyde. ([0047], page 5).
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art before the effective filing date of the rejected claims to combine the teachings of prior art to modify the method of Kang et al by crosslinking gelatin nanoparticles with glutaraldehyde as taught by Skardal et al as instantly claimed, with a reasonable expectation of success. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would have been motivated to do so because Skardal et al teach FIG . 5 that shows construct prepared with a gelatin /GNP bioink that exhibits greatly improved printability ([0015], page 2), and FIG . 6 shows a construct prepared with a bioink including hyaluronic acid , collagen , and gelatin nanoparticles, and the construct was able to be precisely printed and had sufficient strength in that the structure did not collapse or sag, even when tilted sideways” ([0016], page 2). One of ordinary skill in the art would have had a reasonable expectation of success in doing so because Skardal et al were successful in preparation of gelatin bioink to generate constructs that exhibits greatly improved printability and have sufficient strength with detailed instruction and working examples.
Regarding to claim 1, the claimed: floating culturing the cured product, Kang et al teach culturing the artificial tissue in a culture medium (see claim 16, page 9).
However, Kang et al do not teach floating culturing the cured product. McCormack et al cure the deficiency.
McCormack et al teach 3D printing in suspension baths: keeping the promises of bioprinting afloat (title). 3D printing in suspension media unlocks the full potential of extrusion-based 3D printers by providing a strategy for fabricating non–self-supporting structures from water-rich, low-viscosity bioinks (see page 584 in the Highlights box). The inclusion of cells throughout a suspension medium conjures up the idea of the medium acting as a platform to position cells in 3D space, as well as providing a bulk matrix fulfilling some of the functions of a native ECM. The noticeable advantage of suspension media being leveraged in this manner is linked to the opportunity to fabricate larger 3D tissue constructs in a shorter time, increasing the throughput of 3D bioprinting (Page 587 last para. to first para. in page 588).
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art before the effective filing date of the rejected claims to combine the teachings of prior art to modify the method of Kang et al by floating culturing cells in a suspension medium as taught by McCormack et al as instantly claimed, with a reasonable expectation of success. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would have been motivated to do so because McCormack et al provide explicit advantage of inclusion of cells in suspension media is linked to the opportunity to fabricate larger 3D tissue constructs in a shorter time, increasing the throughput of 3D bioprinting (Page 587 last para. to first para. in page 588). Also, suspension media hold unique traits that are responsible for their ability to suspend and completely encapsulate printed material. Suspension media exhibit solid-like characteristics in the absence of an applied stress or at very low stresses such as those induced by gravity. As a result, printing of soft materials or low-viscosity fluids, often containing a high-water content, is feasible with the use of suspension media (Page 585, 2nd para.). One of ordinary skill in the art would have had a reasonable expectation of success in doing so because McCormack et al provide detailed instructions and working examples for using suspension media for tissue engineering.
Regarding to claim 2, Kang et al teaches wherein the cells are selected from the group consisting of stem cells, osteoblasts, myoblasts, tenocytes (e.g., claim 8, Page 8).
Regarding to claim 3, Kang et al teaches (a) preparing a hydrogel, and (c) soling the hydrogel obtained in step (a) at 30 to 40 oC (e.g., claim 11, Page 9). The bio-ink composition according to claim 1, wherein the hydrogel is in a sol state at 30 to 40 oC and gel-state at 4 to 25 oC (e.g., claim 6, Page 8).
Claims 4 is rejected under 35 U.S.C. 103 as being unpatentable over Kang et al (KR 20180125776 A, Date Published: 2018-11-26) in view of Skardal et al (Pub. No.: US 2020/0108172 A1, Pub. Date : Apr. 9, 2020) and McCormack et al (Trends in Biotechnology, June 2020, Vol. 38, No. 6; Doi: 10.1016/j.tibtech.2019.12.020) as applied to claims 1, 2, 3 above, and further in view of Compaan et al. (Pub. No.: US 2020/0189182 Al, Pub. Date: Jun. 18, 2020).
The teachings of the combined references above are incorporated herein in their entirety. Kang et al teaches the hydrogel comprises at least one selected from the group consisting of gelatin, hyaluronic acid, dextran, and collagen (e.g., claim 12, page 9). However, the above references al do not specifically teach gellan gum. Compaan et al cure the deficiency.
Regarding to claim 4, Compaan et al teach “fabricating tissue constructs….. The hydrogel precursor can include at least one of gellan and gelatin. Cross-linking can be carried out chemically, thermally, enzymatically, or physically.” (Abstract). The gelatin-gellan matrix bath material was prepared by dissolving 3% w/v gelatin and 6.8 mM CaC12 (0.1 % w/v CaCl2 .2H20) as needed in jammed 0.5% w/v gellan microgels, then adding transglutaminase. Gellan microgel dispersions were prepared by dispersing the appropriate mass of low acyl gellan (Kelcogel F low acyl gellan gum, Modernist Pantry, York, Me.) ([0088], page 12). Also, Compaan et al teach “…. unmodified gelatin may be used. In some embodiments, the continuous phase can be cross-linked using an enzyme, such as but not limited to transglutaminase (TG), to form stable constructs after printing is complete” ([0056], page 6).
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art before the effective filing date of the rejected claims to combine the teachings of prior art to modify the method of the above combined references by further using gellan gum in fabrication of hydrogels for bioprinting as taught by Compaan et al as instantly claimed, with a reasonable expectation of success. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would have been motivated to do so because Compaan et al teach “The microgel composite matrix bath-based embedded printing approach therefore simultaneously addresses two major concerns in tissue engineering: achieving physiological cell density and perfusing engineered constructs. The matrix bath material design intrinsically elevates the local cell density by a factor of approximately 10, assuming that the microgels occupy 90% of the construct volume. In addition, the ability to print channels within bulk constructs enables fabrication of large-scale perfusable tissue analogues using readily available materials and equipment.” ([0038], page 4). Compaan et al also provide explicit advantage of improving cell survival within bulk cellular constructs ([0081], page 11). One of ordinary skill in the art would have had a reasonable expectation of success in doing so because Compaan et al were successful in generation of hydrogels with addition of gellan gum with detailed instruction and working examples.
Claims 5, 8 are rejected under 35 U.S.C. 103 as being unpatentable over Kang et al (KR 20180125776 A, Date Published: 2018-11-26) in view of Skardal et al (Pub. No.: US 2020/0108172 A1, Pub. Date : Apr. 9, 2020) and McCormack et al (Trends in Biotechnology, June 2020, Vol. 38, No. 6; Doi: 10.1016/j.tibtech.2019.12.020) as applied to claims 1, 2, 3 above, and further in view of Ahn et al (Pub. No.: US 2019/0142995 A1, Pub . Date: May 16 , 2019).
The teachings of the combined references are as described above and are incorporated herein in their entirety. The combined references do not specifically teach the bio-ink is outputted at a pressure of 30 kPa to 100 kPa from a nozzle of 20 gauge(G) to 30 gauge (G) and culturing the culture using a differentiation induction medium. However, Ahn et al cures the deficiency.
Regarding to claim 5, Ahn et al teaches that the ink of the present invention is a hydrogel ([0056], page 4). Ahn et al teaches that for printing by extruding the ink, the pressure which is applied into the ink receiving member may be different depending on the concentration of the ink and the size of the nozzle , but it may be for example, 0.1 to 700 kPa, 1 to 500 kPa or 1 to 700 kPa. ([0034], page 3). Ahn et al teaches that the printing of hydrogels with nozzle sizes of 18 , 20 , 22 , 25 and 27 Gauge was confirmed with a confocal microscope ([0102], page 8).
Regarding to claim 8, McCormack et al teach 3D printing in suspension baths: keeping the promises of bioprinting afloat (title). McCormack et al also teach the inclusion of cells throughout a suspension medium, and advantage of suspension media being linked to the opportunity to fabricate larger 3D tissue constructs in a shorter time, increasing the throughput of 3D bioprinting (Page 587 last para. to first para. in page 588). Additionally, Ahn et al teaches that the cell may be cultured with a cell differentiation material which induces differentiation of cell according to a desired cell line. For example, a stem cell produces a certain range of cell type, by contacting a differentiation medium and being incubated. Multiple types of differentiation media are suitable ([0075], page 6). Thus, it is a matter of choice for a person of ordinary skill in the art before the effective filing date of the rejected claims to perform floating culturing using a differentiation induction medium.
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art before the effective filing date of the rejected claims to combine the teachings of prior art to modify the method of the above references by using pressure of 0.1 to 700 kPa for nozzle sizes of 18 , 20 , 22 , 25 and 27 Gauge and differentiation medium for cell culturing as taught by Ahn et al as instantly claimed, with a reasonable expectation of success. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would have been motivated to do so because Ahn et al provide explicit advantage that a printed product having various cross-sectional patterns can be prepared by printing with high precision and resolution with a three-dimensional method, and when comprising a cell in the biological tissue, a desired shape can be heterogeneously printed and the shear stress on the cell can be largely decreased ([0022], page 2). One of ordinary skill in the art would have had a reasonable expectation of success in doing so because Ahn et al provide detailed instruction with working examples and data to support their proof of principle for using three -dimensional printing with the filled bio-ink.
Response to Arguments
Applicant's arguments filed 09-11-2025 have been fully considered but they are not persuasive. In view of Applicants' amendment of base claim 1, the previous rejections of claims are withdrawn as stated above. Applicants' arguments with respect to the withdrawn rejections and references are thereby rendered moot. The new rejections necessitated by amendments over new cited prior art references of record are as described above and will not be addressed herein.
Conclusion
No claim is allowed.
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/KHOA NHAT TRAN/Examiner, Art Unit 1632
/PETER PARAS JR/Supervisory Patent Examiner, Art Unit 1632